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T and B lymphocytes in human colostrum
Abstract
Studies of human colostral fluid lymphocytes indicated that approximately 50% were T-cells and 34% were B-cells. The remaining lymphocytes could not be definitely classified as belonging to either category. Half of the colostral B lymphocytes were cells bearing surface IgA. Definite stimulation of colostral lymphocytes was recorded using phytohemagglutinin and pokeweed mitogen; however, considerable differences in response to plant mitogens between peripheral blood and colostral lymphocytes were recorded. Human colostrum contained a factor capable of inhibiting lymphocyte PHA response. T-cells present in human colostrum may be capable of transferring important mediator molecules such as transfer factor to newborn infants.
... Various investigators have established the presence of significant proportions of both T and B lymphocytes in human colostrum and milk (5) and have demonstrated their capacity to respond to mitogens in vitro (2,5), synthesize IgA antibody (6), and display in vitro parameters of the delayed hypersensitivities of the donor (1,2). ...
... Various investigators have established the presence of significant proportions of both T and B lymphocytes in human colostrum and milk (5) and have demonstrated their capacity to respond to mitogens in vitro (2,5), synthesize IgA antibody (6), and display in vitro parameters of the delayed hypersensitivities of the donor (1,2). ...
... Although Diaz-Jouanen and Williams (5) have suggested that human cell-free milk, when present in relatively large amounts throughout the entire culture period, will inhibit PHA-induced proliferation, our data indicate that this effect is not evident if the lymphocytes are first washed free of soluble milk components. ...
Human milk lymphocytes (ML) can be partially purified and propagated in vitro as a means of assessing their immunological function. When exposed to a variety of stimuli known to activate T lymphocytes, ML respond in a unique manner that indicates a selected population of immunocompetent cells. ML are hyporesponsive to to nonspecific mitogens and respond in a reduced manner to histocompatibility antigens on allogeneic cells. In most cases, they are completely unresponsive to C. albicans although blood lymphocytes from the same patients respond to the antigen. The Kl capsular antigen of E. coli induces significant proliferation in lymphocytes obtained from milk, but fails to stimulate blood lymphocytes. This dichotomy of reactivity does not appear to result from suppressive factors or cells in milk or insufficient adherent cell function. Rather it appears to reflect the accumulation of particular lymphocyte clones in the breast and the local nature of mammary tissue immunity at the T-lymphocyte level.
... It includes cellular components such as T and B lymphocytes, macrophages, and other mononuclear cells. The colostral cells are immunologically reactive and elicit cell-mediated immunity and synthesize immunoglobulins, predominantly of the IgA class (2)(3)(4). Although appreciable amounts of 7S (serum type) IgA, IgM, and IgG classes of immunoglobulin are found in the colostrum and milk (5), bulk of the immunoglobulin in the colostrum is secretory (llS) type of IgA. ...
... Lymphoproliferative response. The in vitro lymphoproliferative response to phytohemagglutinin (PHA) and purified protein derivative (PPD) in Group 2 subjects was evaluated by the methods described earlier (2,3,8). The optimal dose of PHA and PPD employed was based on individual dose-response curves established previously (8). ...
A group of formula-fed infants were administered a single feed of poliovirus IgA antibody-rich human colostrum 18 to 72 hr after birth. Subsequently, the presence of IgG, IgA, and IgM immunoglobulin and poliovirus antibody activity was determined in serial serum and fecal samples of the neonates. Absorption of IgA immunoglobulin from the colostrum to the circulation was observed in three infants who were fed with colostrum between 18 and 24 hr after birth. Another group of infants of tuberculin-positive mothers who were being breast fed by their own mothers were followed for the development of in vitro correlates of cell-mediated immunity against tuberculin after prolonged breast feeding. Tuberculin-specific proliferative response was observed in the peripheral blood lymphocytes of two neonates after 5 weeks of breast feeding. The responses were undetectable after 12 weeks, although the infants continued to breast feed. No tuberculin reactivity was observed in the cord lymphocytes. These observations suggest uptake of IgA immunoglobulin and components of cellular immunity in the intestine during the immediate neonatal period.
... It is known that different factors modulate immune responses, but in the digestive tract breast milk (BM) may play an important role in the maturation of the immune response, mainly in relation to the gut. In this sense, it has been demonstrated that besides nutrients and antibodies, BM contains cells 5,6 , hormones, cytokines, and growth factors 7,8 as a proline-rich protein 9 and an epidermal growth factor (EGF) 10 . All of these biologically active factors not only facilitate the development of essential digestive functions but also regulate the maturation of the intestinal mucosal barrier. ...
At birth, mammals make the transition from aseptic surroundings to a pathogen-filled environment, it is known that different factors modulate immune responses, but in the digestive tract breast milk (BM) may play an important role in the maturation of the immune response, mainly in relation to the gut. In this sense, it has been demonstrated that besides nutrients and antibodies, BM contains cells, hormones, cytokines, and growth factors. The objective of this work was to evaluate the importance of BM in the development of both the piglet and the intestinal microbiota. For this, two groups of piglets were formed; one suckled with the saw and the other suckled with BM substitute. Stool samples were taken for bacterial identification, this was done through the VITEK-system, and subsequently, the serotype was identified. The results showed that the piglets suckled with BM substitute 18 (100%) presented diarrheal episodes from the 2 nd day, of this group two piglets presented bloody diarrhea, and the group fed with the mother only 7 (38.8%) had diarrheic episodes from the 5 th day. All isolates 285 were Escherichia coli. The largest number of isolates obtained of piglets suckled with BM belonged to serotype O103:H28 and largest number of isolates obtained of piglets suckled with milk substitute belonged to serotype O166:H21. Serotypes belonged to bloody diarrhea were O103:H28 and O166:H21. The present work showed the importance of BM both in the body development of the piglet and in the protection against infections.
... Our demonstration that B lymphoblasts committed to IgA synthesis have the potential to leave GALT and home to the mammary gland does not exclude the possibility that T cells derived from GALT can follow a similar pathway. This idea is supported by the finding that T lymphocytes in milk respond selectively to enteric antigens (36,37). The possibility should be entertained that GALT-derived T cells play a role in development of the secretory IgA system in the mammary gland. ...
Lymphoblasts from the mesenteric lymph nodes (MN) of mice home to the mammary glands of syngeneic recipients late in pregnancy and during lactation, and within hours of transfer most can be shown to contain IgA. Homing does not occur in virgins, in early pregnancy, or after weaning. Homing MN lymphoblasts are sensitive to antiserum to IgA plus complement, but not to other class-specific antisera. Thus, lymphoblasts in MN with the potential to home to the mammary gland are already committed to IgA synthesis and bear surface IgA before reaching their destination. These results explain observations, made by others, of specific IgA antibodies and IgA plasma cells in milk and colostrum after oral immunization. Under natural conditions it is likely that IgA precursor cells, after stimulation in the gut-associated lymphoid tissue by intestinal antigens, migrate to the mammary gland where they secrete antibodies which constitute an important defense mechanism of the newborn. In the absence of lactation, these cells probably form part of the normal traffic to the lamina propria of the small intestine.
... There was a transient increase in IEL in postnatal week 1 in rats, which was suspected to represent a population of maternal lymphocytes from the colostrum (milk lymphocytes; Lyscom and Brueton 1983). This maternal lymphocyte population is known to accumulate in the colostrum of humans (Diaz Jouanen and Williams 1974;Richie et al. 1982) and have been shown to cross the mucosal barrier of the intestine of rats ( Seelig and Billingham 1981). PPs developed in advance of the diffuse IEL population in the present studies. ...
The immune system of the rat undergoes substantial functional and morphological development during the postnatal period. Some aspects of this development are genetically predetermined, while other aspects depend on environmental influences. Detailed information on postnatal development is important in the interpretation of histopathologic findings in juvenile toxicology and pubertal assay studies, as well as other studies conducted in juvenile rats. Studies were conducted to provide detailed characterization of histologic features of the major functional compartments of immune system organs in male and female Sprague-Dawley rats at weekly intervals from the day of birth through postnatal day (PND) 42. Maturation of the individual immune system organs occurred across a range of ages, with histologic maturation of T-cell-related compartments typically occurring prior to maturation of B-cell-related compartments. The sequence of histologic maturation was bone marrow and thymus on PND 14, mesenteric lymph node on PND 21, Peyer's patches and bronchus-associated lymphoid tissue on PND 28, mandibular lymph node, nasopharynx-associated lymphoid tissue, and diffuse mucosal mononuclear cell population of small intestine on PND 35, and spleen on PND 42. An estimation of functional maturation can be made based on the morphological indications of maturity of each compartment of immune system organs, but histologic indications of maturity do not confirm functional immunocompetence.
© 2015 by The Author(s).
... Like the Env-reactive antibodies isolated from colostrum, all clonally related antibodies were IgG1 isotype. This finding was surprising given the previously reported prevalence of IgA-secreting cells in colostrum 7,11,25 and suggests that class switching to IgA is not required for B cells to home to the lactating mammary gland. ...
A successful HIV-1 vaccine must elicit immune responses that impede mucosal virus transmission, though functional roles of protective HIV-1 Envelope (Env)-specific mucosal antibodies remain unclear. Colostrum is a rich source of readily accessible mucosal B cells that may help define the mucosal antibody response contributing to prevention of postnatal HIV-1 transmission. To examine the HIV-1 Env-specific colostrum B-cell repertoire, single B cells were isolated from 17 chronically HIV-infected, lactating women, producing 51 blood and 39 colostrum HIV-1 Env-specific B-cell antibodies. All HIV-1 Env-specific colostrum-derived antibodies were immunoglobulin (Ig)G1 isotype and had mean heavy chain complementarity-determining region 3 (CDR3) lengths and mutation frequencies similar to those isolated from blood. However, variable heavy chain (VH) gene subfamily 1(∼)69 usage was higher among colostrum than blood HIV-1 Env-reactive antibodies (49% vs. 20%, P=0.006, Fisher's exact test). Additionally, more HIV-1 Env-specific colostrum antibodies were gp120 specific than those isolated from blood (44% vs. 16%, P=0.005, Fisher's exact test). One cross-compartment HIV-1 Env-specific clonal B-cell lineage was identified. These unique characteristics of colostrum B-cell antibodies suggest selective homing of HIV-1-specific IgG1-secreting memory B cells to the mammary gland and have implications for targeting mucosal B-cell populations by vaccination.Mucosal Immunology advance online publication, 6 August 2014; doi:10.1038/mi.2014.69.
Human colostrum and milk contain humoral and cellular factors that are important to the immunological protection of the newborn. These humoral components include immunoglobulins of all isotypes, lysozyme, lactoperoxidase, lactoferrin, interferons, and complement factors. The function of colostral cells is not fully understood. It has been suggested that colostral T lymphocytes or their products may confer cell-mediated hypersensitivity to the infant. In addition, T cells from colostrum have been reported to function in cell-mediated immunity, lymphokine production, and to proliferate in response to allogeneic, antigenic, or mitogenic stimulation. Purified cell types are required for the morphological and functional characterization of colostral cells. However, unusual physical characteristics present special problems to the separation and isolation of colostral populations. This chapter discusses the various methods used by the investigators to separate colostral cell populations and to delineate the efficiency and drawbacks of each. Adherence is the most commonly used method for the separation of colostral cell populations. It takes advantage of the ability of polymorphonuclear leukocytes (PMN) and macrophages to adhere to glass or plastic surfaces.
Knowledge of the histogenesis of immune system organs is important in the detection and interpretation of histopathologic alterations in nonclinical toxicology studies, particularly studies that are conducted in juvenile animals (exposed either in utero or postnatally). In addition to the direct, indirect, nutrition-, and stress-related histologic alterations that are encountered in immune system organs in nonjuvenile toxicology studies, in juvenile toxicology studies the pathologist and toxicologist must also be concerned with the normal postnatal development and the potential for xenobiotic-associated influences on those processes. In addition to genetically determined, age-related differences, the histomorphology of the secondary immune system organs is heavily influenced by environmental factors, thus test article-related influences on nonimmune organs or functions, for example, the microbiome of the gastrointestinal tract, may have secondary effects on immune system organs. When confronted with these challenges, it is important that pathologists and toxicologists have an understanding of the embryological and postnatal development of immune system organs. Given the emphasis on histopathologic evaluation in detection of xenobiotic-associated influences, this article is focused on histomorphological rather than functional alterations. The laboratory rat is commonly used in nonclinical toxicology studies, thus the rat is used as the major example of processes that occur during the development of immune system organs.
In addition to the direct, indirect, nutrition- and stress-related histologic alterations encountered in immune system organs of adult animals in toxicology studies, in juvenile toxicology studies the pathologist and toxicologist must also be concerned with the influence of normal postnatal development on the histologic appearance and functional attributes of immune system tissues. In addition to direct age-related differences in histomorphology, the postnatal development of the immune system is partially controlled by environmental factors, thus test article-related influences on non-immune organs or functions, e.g., the microbiome of the gastrointestinal tract, may have secondary effects on immune system organs. It is important that pathologists and toxicologists have a thorough understanding of the normal embryological and postnatal development of immune system organs in order to prevent misinterpretation of these normal changes as xenobiotic-associated. This chapter is focused on histomorphological rather than physiological alterations, though the intent certainly is not to dismiss the importance of traditional immunological assays in the detection of xenobiotic-associated influences. The laboratory rat is commonly used in nonclinical toxicology studies, thus the rat is used as the major example of the processes that occur during the postnatal development of immune system organs. An attempt is made to provide cursory information relating developmental and homeostatic changes in the rat as compared to those seen in other commonly used lab animals and humans, but a thorough presentation of these changes in humans and all animal species is beyond the practical scope of this Chapter.
During the last 15 years, progress in understanding the structure and functions of lymphoid organs, cells, and molecules has provided us with a better understanding of their role in various immunological phenomena. This has brought us to a point of clinical application. Animal experiments and studies of experiments of nature in humans have led to the discovery of a two-component concept of immunity (Cooper et al., 1965). Figure 1 depicts our current concept of this two-cell system of immunity. It is now clear that cells originating from the hemangioblasts of yolk sac develop into mature hematopoietic cells. It seems likely that cells of the yolk sac contain the precursors of all the major hematopoietic elements. During development, these pluripotent stem cells migrate to the fetal liver, where they not only can give rise to an additional population of stem cells but also may be induced to differentiate along several additional lines.
Four major aspects of the immune-inflammatory response are considered, including humoral immunity, cell-mediated immunity, phagocytic function and humoral mediators, particularly complement. This is followed by a discussion of the role of human milk in enhancement of neonatal host defenses (an extremely active area of current immunologic research) and a clinical summary.
Colostrum contains large numbers of cells and humoral defense factors, that play an important role for the protection of the neonate. While colostral cells have been characterized by immunohistochemical criteria, the fine structure of these cells has not been studied. In this study, colostral cells were examined using a combination of electron microscopy and histochemical techniques. Colostral neutrophils and macrophages were found to contain numerous phagocytic vacuoles that stained positively for acid phosphatase. These suggest that these cells actively phagotized various substances from the environment and that phagocytic vacuoles were secondary lysosomes. Distribution of the peroxidase activity in colostral macrophages suggested that they were derived from the circulation. By SEM, neutrophils were found to be circular or spherical cells and showed a rough surface with several blebs in limited areas, while macrophages ranged in shape from spherical to polyhedral and demonstrated numerous ridges or ruffles over the entire cell surface. Lymphocytes were small spherical cells with a smooth surface and a few villous processes. Macrophages were distinguishable from neutrophils and lymphocytes by size and surface structure.
The most important factors that determine the occurrence and severity of diarrheal disease include a safe environment, consisting of an efficient water source and sewage removal system; adequate personal and food hygiene; and good standards of nutrition. Despite the presence of these factors, the human body is exposed continually to pathogenic and nonpathogenic microorganisms, toxic substances, and chemicals which would be damaging or fatal if not efficiently excluded by the gastrointestinal tract. Once organisms are swallowed, the occurrence of a diarrheal syndrome will depend upon the pathogenic potential of the agent (virulence), the number of viable cells or viruses ingested, and the status of host defense factors.
This chapter discusses the acquisition of immunity by the neonate. During mammalian evolution a number of different strategies for the provision of maternal antibody to the neonate have arisen. However, a common theme emerges in that, despite wide differences in pre- and postnatal immune transfer and in immunoglobulin isotype contribution, the maternal contribution is designed to protect her mature offspring from both systemic and mucosal infections without compromising the future immune responsiveness of the neonate. The routine suppression of neonatal immune responses or the tolerance of the neonate to potentially dangerous antigens by maternal elements would be biologically unacceptable. The passive protective postnatal contribution is mediated by the maternal lacteal secretions; colostrum and milk. Such a system is necessary for the optimum development of the neonate is illustrated by many studies. The chapter also discusses the duration and functionality of specific immunity in the newborn mediated by maternal colostrum and milk. Such specific immune reactivity may be separated broadly into two areas; passive antibody-mediated immunity and the more nebulous passive cell-mediated immunity. The chapter also examines the integration of the developing immune system of the neonate with declining levels of passive maternal immunity.
Human milk contains antibodies of the secretory IgA (SIgA) type directed against a variety of enterobacteria, viruses and food proteins. The appearance of these milk antibodies with specificity against antigens of intestinal origin is difficult to explain, unless there is a close link between the immune response in the mammary gland and intestinal exposure. There is some evidence which suggests that antigens in the intestine may stimulate lymphoid cells from the Peyer’s patches which are committed to IgA synthesis to home to various exocrine glands, including the mammary gland. As a consequence of the enteromammaric Zink, the breast-fed baby is supplied with SIgA milk antibodies against micro-organisms which are common in the environment. These antibodies have the capacity to prevent bacterial attachment to mucous membranes which is the initial step in most infections. Determination of milk antibodies also provides epidemiological information concerning intestinal pathogens and can be used to reflect mucosal immunity in response to various vaccinations.
Mucosal immunity is charascterized predominantly by a secretory antibody response, involving immunoglobulins of the IgA class. They provide an immunological barrier to foreign matter by preventing absorption of such materials into the mucosal epithelium and penetration into the body. The humoral responses in nonmucosal sites are largely of the IgG class. The cell-mediated immune mechanism is also distinctive in mucosal and nonmucosal lymphoid organs. During the past decade, significant advances have been made in understanding the mucosal immune system. Ontological and physiological relationships between immunocompetent cells in mucosal and nonmucosal sites have been characterized, and the postulation of a common mucosal immune system1,2 involving the intestinal tract, biliary tract, mammary glands, salivary glands, lacrimal glands, and female genital tract has led to new insights.
The effects of pasteurization, lyophilization, and freezing on immunoglobulin and lymphocytes in human milk was studied. We found a significant decrease in total lymphocyte count after all three processing methods. Pasteurization and lyophilization caused a significant decrease in immunoglobulin concentration and in specific antibody titer to Escherichi coli . Freezing specimens up to four weeks resulted in no significant alteration of IgA content or in E. coli antibody titer. Since a major advantage of human milk is the transference of passive local immunity, these quantitative changes may have clinical importance.
Resistance to infection and protection from allergic disorders (or atopy) are probably the most frequently cited immunologic benefits of breast-feeding. In this chapter, some of the immunologic constituents of breast milk are described and the mechanisms of their possible beneficial effects discussed. In order to provide a background for this discussion, the basic effector mechanisms involved in immune responses are summarized in this section. Special attention has been given to the immune processes occuring at mucosal and secretory surfaces such as the intestinal and mammary epithelium.
Human κ-caseinoglycopeptides were prepared from acid human casein fractions digested with rennin or pepsin. The glycopeptides inhibited the proliferative response of mouse spleen and rabbit Peyer's patch cells induced by lipopolysaccharide, pokeweed mitogen, phytohaemagglutinin and concanavalin A stimulation, and suppressed immunoglobulin production in mouse spleen cell cultures with or without lipopolysaccharide. Moreover, the glycopeptides retained cyotoxic activities towards mouse spleen cells and Jurkat cells derived from human acute leukemic T-lymphocytes. An estimation of the 3′-OH end of DNA (TUNEL staining) in Jurkat cells cultured with glycopeptide and agarose gel electrophoresis of DNA extracted from cultured Jurkat cells showed DNA cleavage into oligonucleosome-sized fragments. These results indicate that human κ-caseinoglycopeptide possesses not only suppressing activities towards lymphocyte proliferation and immunoglobulin production but also an apoptosis-inducing ability towards some lymphocytes including tumor lymphocytes.
A most important adaptation of the G.I. tract of the newborn to the extrauterine environment is its development of a mucosal barrier against the penetration of bacteria viruses and other environmental antigens i.e food proteins. The mechanisms of host defence at mucosal sites both im-mulogic and nonimmunologic are underdeveloped during the neonatal period and more so in prematures neonates (1).
Infectious diseases take the lives of many infants and young children in developing countries. Furthermore, the adverse effects of infection linger even among the survivors who may suffer growth faltering and its associated risks, including, in the short term, increased susceptibility to infection (Tomkins and Watson, 1989) and, in the long term, decreased life-long productivity (Gopalan, 1988). Because of the devastating effects of infectious disease, much research has dealt with possible means of prevention of infection, including immunization and improved sanitation, water supplies and personal hygiene. One factor which appears to have an important influence on an infant’s risk of and response to an infection is the infant’s diet. Infant feeding practices and their relation to infectious disease are the subject of this review, with particular emphasis on the role of breast feeding but also with some discussion of weaning foods.
The mammary gland (MG) lacks a mucosa but is part of the mucosal immune system because of its role in passive mucosal immunity. The MG is not an inductive site for mucosal immunity. Rather, synthesis of immunoglobulin (Ig)A by plasma cells stimulated at distal inductive sites dominate in the milk of rodents, humans, and swine whereas IgG1 derived from serum predominates in ruminants. Despite the considerable biodiversity in the role of the MG, IgG passively transfers the maternal systemic immunological experience whereas IgA transfers the mucosal immunological experience. Although passive antibodies are protective, they and other lacteal constituents can be immunoregulatory. Immune protection of the MG largely depends on the innate immune system; the monocytes-macrophages group together with intraepithelial lymphocytes is dominant in the healthy gland. An increase in somatic cells (neutrophils) and various interleukins signal infection (mastitis) and a local immune response in the MG. The major role of the MG to mucosal immunity is the passive immunity supplied to the suckling neonate.
This chapter focuses on immunoglobulins and immunocytes in milk. The chapter outlines the mammary gland anatomy and milk biochemistry. Under the influence of the hormones of pregnancy and lactation such as prolactin, progesterone, and local growth factors—the narrow ducts bud alveoli and parenchymal tissue progressively replaces the fat pads of the undeveloped mammary gland (MG). During lactation, leukocytes are confined to clusters between distended alveoli. Many milk cells are associated with immunoglobulins, including T cells. Much of the surface immunoglobulin expression on milk cells reflect immunoglobulins bound to Fc receptors, which decorate neutrophils and monocytes in all species. Cells from blood enter the MG, transit the alveolar epithelium, and enter the milk. The higher expression of CD45RO on both milk CD4 and CD8 cells also resembles that of T cells in the gut and buccal mucosa. Some differences between blood and milk lymphocytes could be microenvironmental. Experimental studies suggest that milk lymphocytes are less responsive to mitogens and that colostrum can inhibit the mitogenic response of peripheral blood lymphocytes.
During recent years much information has accrued concerning factors in human milk which may help the breast-fed infant in its defense against infections. Some of these factors, especially those with specific immune functions, will be briefly reviewed.
The ability of colostral lymphocytes to respond to pokeweed mitogen, phytohemagglutinin, or Epstein-Barr virus was examined. None of these mitogens induced colostral cells to differentiate into immunoglobulin-containing cells, either in the absence or in the presence of mitomycin C-treated mononuclear cells or T-cell-enriched populations from peripheral blood. Cocultivation of mononuclear cells from the peripheral blood of normal adults with mitomycin C-treated colostral cells resulted in a marked suppression of the generation of immunoglobulin-containing cells in response to pokeweed mitogen. The inhibitory effect was seen in peripheral blood mono-nuclear cell:colostral cell ratios of 1:1, 5:1, and 10:1. However, colostral cells had little effect on the ability of peripheral blood lymphocytes to proliferate in response to phytohemagglutinin or to allogeneic stimulation.
The phagocytic properties of sow milk cells and their capacity for intracellular killing of E. coli cells were investigated. The phagocytic activity of the cells were compared with those of blood leukocytes. It was found that the milk cells phagocytosed E. coli to a lower degree than did blood leukocytes. The milk phagocytes are defective in intracellular killing of phagocytosed bacteria. Since the cells poorly reduce nitroblue tetrazolium it is possible that the lack of intracellular killing results from impaired aerobic metabolism of the milk phagocytes. Sow milk had opsonizing properties but they were poorer than those of serum. Milk cells survived better in an environment with an acidic reaction than an alkaline one.
Zusammenfassung
Zellen im Milchdrüsensekret der Sau III. Einige Eigenschaften der phagozytierenden Zellen
Es wurden einige Phagozytose‐Eigenschaften der Zellen im Milchdrüsensekret der Sau sowie die Fähigkeit zur intrazellulären Abtötung von E. coli ‐Zellen untersucht. Die Phagozytoseaktivität der Zellen wurde mit derjenigen von Blut‐Leukozyten verglichen. Milchzellen phagozytierten E. coli in geringerem Umfang als Blut‐Leukozyten. Bei den Phygozyten der Milch ist zudem die Fähigkeit zur Abtötung der phagozytierten Bakterien gestört. Da die Zellen Nitroblau‐Tetrazolium schlecht reduzierten, dürfte die mangelhafte intrazelluläre Abtötung auf einer Beeinträchtigung des aeroben Stoffwechsels der Milch‐Phagozyten beruhen. Sauenmilch wies opsomierende Eigenschaften auf, die jedoch weniger ausgeprägt waren als diejenigen des Serums. Milchzellen überlebten in einem sauren Medium besser als in einem alkalischen Medium.
Se exponen en forma de revisión los factores inmunológicos presentes en la leche materna que protegen al niño durante los primeros años de vida. Se hace referencia a las ventajas clásicas que muestra la leche materna y al limitado conocimiento existente entre los trabajadores de la salud, de los elementos inmunológicos presentes en la leche materna que actúan como la primera vacuna que recibe el niño después del nacimiento. Se describe el eje entero-bronco-mamario, así como el sistema común de defensa de las mucosas. Se presentan en detalles los componentes celulares y humorales inmunoprotectores de la leche materna, y se enfatiza en su elevado contenido de inmunoglobulina A secretora. Se enumeran los anticuerpos bacterianos y virales, así como los factores antibacterianos, antivirales y antiprotozoarios, no anticuerpos presentes en la leche materna. Se desarrollan los aspectos más relevantes de estos factores, con destaque de la lactoferrina, la lisozima, el factor bífido, la lipasa estimulada por sales biliares, etcétera. Se hace referencia a los linfocitos "B" y "T", a los macrófagos y los leucocitos polimorfonucleares. Se subraya que los médicos y enfermeras deben conocer estos elementos, para estar convencidos de las bondades de la leche materna y convertirse en los mejores promotores de su uso.
Milk contains a number of immunoprotective and antimicrobial agents, aimed at the protection of the newborn animal. It contains specific immunoglobulins as well as non‐specific bactericidal factors. It mimics the defence mechanisms present in the blood stream.The immunoglobulin composition is related to the need for immune support in the young animal, which differs between e.g. primates and ruminants.The non‐specific factors present in milk of different species are mostly the same in different species, although the amount and their importance can vary widely. There are three factors that dominate: Lactoperoxidase, lactoferrin and lysozyme. They are all very basic proteins what renders them special physico‐chemical properties.These three non‐specific factors interact with each other and with e.g. sIgA. How these aggregations occur in vivo and whether they regulate the antimicrobial properties in milk ought to be further investigated.
The distribution of T-lymphocytes and other cells and proliferative activity to a variety of bacterial and viral antigens was studied in the colostrum and milk and peripheral blood of postpartum subjects at various times after the onset of lactation. The techniques of E-rosette formation and in vitro lymphoproliferative responses were employed to study the responses in colostrum and milk and peripheral blood lymphocytes. The highest number of total cells (105-107/ml) and of E-rosetting lymphocytes (104-105/ml) were observed in the colostrum on the first postpartum day. Subsequently, the concentration of cellular components decreased sharply and by the fourth postpartum day the number of E-rosette forming cells in milk had dropped by about 10-fold, and the number of E-rosette forming cells 180 days postpartum was about 103/ml. No change was observed in the cell counts in the peripheral blood. The proliferative responses to phytohemagglutinin, concanavalin A, streptokinase, candida, rubella, cytomegalovirus, and mumps virus antigens were frequently detected in the colostrum lymphocytes. However, the activity in colostrum lymphocytes appeared to be independent of the activity in the peripheral blood lymphocytes. Little or no responses were observed in the milk collected 15 to 180 days postpartum, in spite of an adequate number of E-rosette forming cells. In addition, colostrum and milk were found to have a profound inhibitory effect on the proliferative responses of perpheral blood as well as on colostrum and milk lymphocytes.
The neonatal period is one of great importance for infant survival in low income countries. Breast‐feeding, particularly feeding of colostrum, is recommended to be initiated as quickly as possible during this period. Much has been written about breast‐feeding patterns in low income countries. Nevertheless, few quantitative descriptions on feeding of the neonate are available. This study describes in detail patterns in the consumption of colostrum, prelacteal feeds and breast‐milk substitutes in the first two days of life. A prospective survey of 3080 infants representative of rural and urban areas in the Metropolitan Cebu region of the Philippines comprises the sample.There was considerable heterogeneity of feeding in this sample, particularly in urban areas. Feeding of prelacteal foods was the dominant pattern in the first day, while exclusive breast‐feeding was the major pattern in the second day. Proportions of women feeding prelacteal foods and breast‐milk substitutes were higher among urban women and those with hospital deliveries than among rural women and those having home deliveries. The amounts of colostrum and other prelacteal feeds consumed varied by feeding pattern.
Forty–three specimens of human milk were obtained from allergic and non–allergic women 3–6 days post–partum and analyzed for their eventual content of eosinophils and metachromatically staining cells. Eosinophils and metachromatically staining cells were identified in all milk samples; however, in lower numbers than in peripheral blood (0.082 to 4. 44% and 0.002 to 0.098% resp. of total cell numbers). Possible contamination of milk fluid with peripheral blood was ruled out. As no mast cells were detected with monoclonal anti–tryptase antibodies, the metachromatically staining cells were basophils. The numbers of basophils and eosinophils in milk were positively correlated to each other (p < 0.05) and to blood eosinophil counts (p0 < 0.001 and p1= 0.01, respectively), suggesting migration of these cells from blood to the mammary glands. Significantly higher numbers of eosinophils, but not of basophils were encountered in milk of allergic women compared to milk of non–allergic mothers (p0 < 0.01 and p1 > 0.05, respectively). As histamine release in the gut increases gut permeability and thereby probably the risk for sensitization to food allergens, basophils in milk might represent an allergy risk factor for the suckling neonate.
The normal immunity of the newborns, or rather the immune deficiencies, are the expression of the development of the immunological systems and functions during intra-uterine life. This development normally takes place in the absence of antigenic stimulation.The development of the immunological system occurs at the 16th week as regards the thymus, the 25th week as regards the spleen, and later still for the lymphatic ganglia. The body of the organ (entoblastic or mesoblastic) is formed before it becomes colonized by lymphocytes of medullary origin. Normally there are no plasmocytes. Differentiation of the lymphocytes (T and B) occurs very early as does the synthesis of the different complement factors. Immunoglobulins are synthesized from the 74th day. The A and B blood group antigens, the antigens of the HLA system are present from the 10th week.Antenatal immunological responses are possible (during the third trimester of fetal life), being expressed by the appearance of specific antibodies (mainly IgM, but also IgE), or of cellular immunity.At birth, the maternal contribution of immunoglobulins makes up for the deficiencies of the newborn. Local immunoglobulins are not present. The humoral immunity of the newborn is passive. Inversely, the newborn can present immediate, active hypersensitivity phenomena.In conclusion, the authors draw attention to the interrelations of the immunological systems of the mother and of the fetus and recall that the control mechanisms prevent reciprocal effects.
Breast milk has replaced proprietary formulas as an important feeding for the premature and sick neonate. Over 30 milk banks have been established in the United States to provide breast milk to high-risk neonates. Planning a milk bank poses many problem as no uniform standards for quality control exist. A format for developing a breast milk bank and the protocol for donor screening, collection techniques, transport and storage of milk are presented. Rationale supporting each section of the protocol is also included.
The IgG, IgA and IgM synthesis by adult peripheral blood and cord blood lymphocytes incubated alone and with pokeweed mitogen was quantitated. The cord blood lymphocytes produced no immunoglobulin even with mitogen stimulation while the adult peripheral blood lymphocytes responded to the mitogen with a significant (p
Characterization of lymphocyte subpopulations in the bovine mammary gland was accomplished using cells obtained from dry secretions. Correlation of cell surface properties with functional capacity was attempted by assaying the ability to form erythrocyte-antibody (EA) rosettes, erythrocyte-antibody-complement (EAC) rosettes, and sheep erythrocyte (E) rosettes and the ability to respond to phytohemagglutinin (PHA), Concanavalin A (Con A), and pokeweed mitogen (PWM) in the lymphocyte stimulation test. Results were compared with those obtained for peripheral blood lymphocytes (PBL) from the same animals. Mammary gland lymphocytes (MGL) formed significantly fewer (p < .01) EA and EAC rosettes, but significantly greater (p < .01) E rosettes compared to PBL. MGL were significantly less responsive (p < .05) to mitogens than were PBL. MGL contained a large proportion of T lymphocytes, which do not respond to T lymphocyte mitogens in culture.
Breastfeeding accounts for up to 40% of all infant human immunodeficiency virus (HIV) type 1 infections in resource-limited settings, where prolonged breastfeeding is the only available and safe infant feeding option [1, 2]. However, most breastfed infants remain uninfected even after prolonged exposure to breast milk [3-5]. The factors in breast milk that protect the majority of breastfed infants of HIV-infected mothers from infection remain largely undetermined. Breast milk contains a multitude of immune parameters, including immunoglobulins, antimicrobial substances, pro-and anti-inflammatory cytokines, and leukocytes [6]. Moreover, breast milk not only provides passive protection, but also can directly modulate the immunological development of the infant [7, 8]. Cell-mediated immunity in breast milk has not been as extensively studied as humoral immunity, described in Chap. 10. There is, however, increasing interest in the role that lymphocytes, macrophages, and other immune cell types play, both in innate and in adaptive breast milk immunity [3].
Breast milk transmission of HIV is a leading cause of infant HIV/AIDS in the developing world. Remarkably, only a small minority of breastfeeding infants born to HIV-infected mothers contract HIV via breast milk exposure, raising the possibility that immune factors in the breast milk confer protection to the infants who remain uninfected. To model HIV-specific immunity in breast milk, lactation was pharmacologically induced in Mamu-A*01(+) female rhesus monkeys. The composition of lymphocyte subsets in hormone-induced lactation breast milk was found to be similar to that in natural lactation breast milk. Hormone-induced lactating monkeys were inoculated i.v. with SIVmac251 and CD8(+) T lymphocytes specific for two immunodominant SIV epitopes, Gag p11C and Tat TL8, and SIV viral load were monitored in peripheral blood and breast milk during acute infection. The breast milk viral load was 1-2 logs lower than plasma viral load through peak and set point of viremia. Surprisingly, whereas the kinetics of the SIV-specific cellular immunity in breast milk mirrored that of the blood, the peak magnitude of the SIV-specific CD8(+) T lymphocyte response in breast milk was more than twice as high as the cellular immune response in the blood. Furthermore, the appearance of the SIV-specific CD8(+) T lymphocyte response in breast milk was associated with a reduction in breast milk viral load, and this response remained higher than that in the blood after viral set point. This robust viral-specific cellular immune response in breast milk may contribute to control of breast milk virus replication.
The possibility that cell-mediated immunity could be acquired by breast-feeding was evaluated in a prospective study of 26 tuberculin positive and 9 negative puerperal mothers and their infants. 13 infants of the positive and all the infants of negative mothers were breast-fed. Tuberculin-reactive T cells were found in colostrum and early milk of most positive but in none of the negative nursing mothers. A significant number (8/13) of infants born to positive mothers had tuberculin-reactive peripheral blood T cells after 4 weeks of breast-feeding compared with bottle-fed infants (1/13) of positive mothers or breast-fed infants (0/9) of negative mothers. Examination of cord blood for tuberculin-reactive T cells provided no significant evidence of transplacental transmission of responsiveness to tuberculin. The results suggest that breast-fed infants may passively acquire T cell responsiveness to a specific antigen by ingestion of breast milk.
Human milk lymphocytes (ML) can be partially purified and propagated in vitro as a means of assessing their immunological function. When exposed to a variety of stimuli known to activate T lymphocytes, ML respond in a unique manner that indicates a selected population of immunocompetent cells. ML are hyporesponsive to to nonspecific mitogens and respond in a reduced manner to histocompatibility antigens on allogeneic cells. In most cases, they are completely unresponsive to C. albicans although blood lymphocytes from the same patients respond to the antigen. The Kl capsular antigen of E. coli induces significant proliferation in lymphocytes obtained from milk, but fails to stimulate blood lymphocytes. This dichotomy of reactivity does not appear to result from suppressive factors or cells in milk or insufficient adherent cell function. Rather it appears to reflect the accumulation of particular lymphocyte clones in the breast and the local nature of mammary tissue immunity at the T-lymphocyte level.
Bovine and canine milk contained lymphocytes that responded to the nonspecific mitogens; phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) and gram negative bacterial lipopolysaccharide (LPS). It also was found that animals specifically sensitized with tuberculin or infected with infectious bovine rhinotracheitis virus (IBR) had antigen sensitive lymphocytes in their milk. In general, the responses of the milk lymphocytes from an individual animal were not identical to responses for the blood lymphocytes. Marked variation was observed in the daily responses of cells from milk samples from different quarters of the same animal, and between animals. The implications of antigen and mitogen responsive milk lymphocytes are discussed, in relation to their possible role in protective immunity.
Human colostrum and milk contains T and B lymphocytes, macrophages and the major classes of immunoglobulins (1). The bulk of the immunoglobulins in colostrum and milk belong to the secretory (11S) IgA class (2,3). It has been suggested that the secretory IgA antibody activity found in the mammary glands and in colostrum is largely derived from the gut associated lymphoid tissue (4). Little or no information is available regarding the distribution of antibody producing cells and the levels of immunoglobulins in colostrum and milk at various times after onset of lactation. Fragmentary data are available on T-lymphocyte function and in vitro correlates of cell-mediated immunity in colostrum and milk (1,4). Little is known about the intestinal absorption of the immunoglobulins and cellular components in colostrum.
Experiments were performed on human B-lymphocytes in paired samples of milk and blood from the same patient to determine the numbers and percentages of lymphocytes with complement receptors, utilizing the erythrocyte-antibody-complement (EAC) rosette assay. Studies on nine paired samples show the numbers, as well as the percentages, of complement-reactive cells (CRCs) in milk are significantly lower than in blood. Milk lymphocytes were cultured in media containing serum for a 24 hour period to determine if the observed lower numbers of CRCs in milk were due to saturation of the lymphocyte receptors with free complement, as well as to determine if the culture of the cells in nutrient media might result in the reformation of shed complement receptors. Similarly, peripheral blood lymphocytes were incubated in milk supernatant to determine if free complement or other factors in milk might interfere with the EAC assay. These studies support previous findings from this laboratory that B-lymphocytes in milk represent a subpopulation of cells different from those in peripheral blood. The paucity of B-lymphocytes bearing complement receptors in milk may be due to the following: (1) B-lymphocytes in milk may be plasmablasts and/or antibody-producing plasma cells--both known to lack complement receptors; (2) selective processes operating at the mammary alveolar epithelium allowing transit of certain subclasses of maternal immunoglobulins may also allow transepithelial passage of B-lymphocytes with surface immunoglobulins, but no complement receptors. These possibilities are currently being studied.
Historically colostrum and milk have been thought to confer immunity on the neonate only by virtue of their immunoglobulin content. Recently we have observed that colostrum also contains viable T lymphocytes capable of expressing cell-mediated immunity in vitro and have employed techniques of lymphocyte culture to elucidate the local nature of mammary tissue immunity at the T-cell level. The results indicate that the activity of colostral lymphocytes appears not to represent the total immunological experience of the mother but that they may contain reactive clones beneficial for the suckling. Colostral immunity appears to depend upon sensitizing events within the intestine and respiratory tract, followed by the migration of lymphoid precursors to the breast, suggesting a relationship between the expression of immunity at various secretory surfaces.
The γ1A present in saliva and colostrum exists largely in the form of higher polymers, the major component of which has a sedimentation coefficient of 11S. The 11S γ1A in these fluids differs from the polymers found in normal and myeloma sera both immunologically and by the fact that their sedimentation coefficients are unaffected by disulfide bond reduction in the absence of urea. However, like other γ-globulins the 11S γ1A molecules consist of multiple polypeptide chains linked by disulfide bonds.
Local synthesis of γ1A in the salivary gland has been shown by fluorescent and autoradiographic studies, although the fraction of the total salivary γ1A which is derived from local production is uncertain. No evidence of transport of intravenously administered I131-labeled 7S γ1A from serum to saliva was obtained.
Immunological specificity has been demonstrated in the salivary and colostral γ1A. Whether that portion of the γ1A which is immunologically specific is a piece incorporated during the local synthesis of γ1A in the gland or is added by the epithelial cell in the process of transport remains to be determined.
Antibody activity (isohemagglutinins) have been demonstrated in saliva and colostrum and have been shown to be of the γ1A-type. In both of these fluids activity is associated primarily with γ1A-polymers of 11S and 18S sizes. There appears to be an immunological system which is characteristic of certain external secretions. Its properties including the local production of a distinctive type of antibody separate it from the "systemic" system responsible for the production of circulating antibody. This system may play a significant role in the body's defense mechanisms against allergens and microorganisms.
Small and medium lymphocytes from the peripheral blood and lymphoid tissues of the rabbit react in suspension with antibodies directed against different immunoglobulin determinants. Through immunofluorescence, it was possible to show that numerous discrete spots on the surface of the positive lymphocytes carry immunoglobulin molecules.
The positive lymphocytes are about one-half of all lymphocytes in the different preparations; thymus lymphocytes are all negative.
With antisera specific for rabbit IgM as well as with antisera directed against allotypic determinants specific for IgM or IgG, it was possible to show that about nine-tenths of the immunoglobulin-positive lymphocytes carry IgM molecules on their surface.
With antisera directed against a- and b-locus determinants, it was also possible to demonstrate that both heavy and light chains were present in the surface immunoglobulins. Furthermore, in animals which were heterozygous at the a or the b locus, it was found that each lymphocyte had immunoglobulins synthesized under the influence of only one of two alleles.
A very small proportion of lymphocytes could be shown to have a specific surface reaction with one antigen (horse ferritin); the proportion of these cells increased very much after immunization.
1. Five patients with congenital or acquired agammaglobulinemia, lacking detectable IgA in serum or saliva, were transfused with 1 to 2 liters of normal plasma. In 2 of these patients IgA was demonstrated in parotid saliva collected after transfusion, but in none of the 5 was salivary IgG or IgM found. This observation indicates the selective transport of IgA into saliva.
2. The observation by others of an immunochemical difference between serum and sahvary IgA globulin was confirmed. In contrast to serum IgA, salivary IgA is attached to a protein having antigenicity which migrates as a gamma1 globulin. We have termed this protein component "transport piece".
3. The transport piece has been found in an unbound form in the saliva of persons completely lacking IgA: agammaglobulinemic patients, ataxia-telangiectasia patients, a healthy person lacking IgA, and a newborn infant. Free transport piece still occurs in the normal child's saliva after IgA production begins. By adulthood there is usually no free transport piece in the saliva.
4. Heat-aggregated salivary IgA, like heat-aggregated serum IgA, does not fix complement.
5. Our findings offer support for the view that there is a distinct local antibody system for the protection of the mucous surfaces.
Immunofluorescent studies using live cells from antibody-forming organs and anti-immunoglobulin antibodies demonstrate two
populations of small lymphocytes which are differentiated by the presence or absence of Ig on their surface membranes. Most
of the lymphocytes with detectable surface Ig appear to derive from cells of the bone marrow, while most of the Ig-negative
lymphocytes derive from the thymus. Thus, adult mice thymectomized, lethally irradiated, and transplanted with bone marrow
cells showed a normal number of lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes. Injection of
thymocytes into these mice did not result in an increase in the number of lymphocytes with surface Ig in spleen and lymph
nodes. Most of the injected thymocytes could be identified by means of histocompatibility markers. Also, the spleen and lymph
nodes of neonatally thymectomized mice contained lymphocytes with surface Ig but were depleted of the Ig-negative lymphocytes.
Attempts were made to identify light chains on thymocytes by a sensitive radioimmunoassay. In some experiments no light chains
were detected and in others a small amount, i.e. no more than 2.6% of the amount present on spleen lymphocytes, could be detected.
Whether these low figures are significant or represent a small amount of serum contamination is not clear as yet.
The distribution, and quantity of immunoglobulins on the surface of lymphocytes has been studied by means of immunofluorescence and a quantitative radio-immunoassay. Surface immunoglobulins were found on approximately 45% of spleen and marrow lymphocytes and 7-14% of lymphocytes from lymph nodes, peripheral blood, and peritoneal exudate. Thymic lymphocytes contained undetectable amounts of immunoglobulin. In the spleen the different immunoglobulins were present in the following order: gammaG2 > gammaG1 > M > gammaA > gammaG3. The surface immunoglobulin was largely removable by brief treatment with trypsin. Quantitative analysis indicated that 50,000-150,000 molecules of immunoglobulin were present on an individual cell. A variety of observations make it likely that this lymphocyte-associated immunoglobulin. is a product of the cell to which it is attached rather than a form of cytophilic antibody.
By using the two criteria (a) high density of immunoglobulin determinants on the cell surface and (b) presence of receptors for C'3 on the cell surface for defining bone marrow-derived lymphocytes, it is indirectly shown that all or at least a major population of human thymus-derived lymphocytes under certain conditions will form nonimmune rosettes with sheep red blood cells (SRBC). Almost all thymocytes tested from two different donors formed rosettes. The SRBC rosettes are not formed by virtue of immunoglobulin receptors and form only around living cells. Positive bivalent ions are required for rosette formation since EDTA will block rosette formation. Sodium iodoacetate will also block rosette formation demonstrating the dependence on an intact glycolytic pathway. Rosette formation is temperature dependent and will not appear at 37 degrees C. Trypsin treatment of lymphocytes will abolish their SRBC-binding ability which cannot be restored by treating them with fresh donor serum or fetal calf serum, but which will reappear after culturing the lymphocytes. It is suggested that these rosettes are formed by a rapidly released or metabolized receptor substance on the living cell surface which behaves as a trypsin-sensitive structure produced by the cells themselves.
Lactoferrin, an iron-binding protein previously shown to occur in many external secretions, is identified as one of the major
proteins present in human and guinea pig neutrophilic polymorphonuclear leukocytes.
The identification of this protein in leukocyte extracts was based upon a comparison of its electrophoretic, antigenic, and
iron-combining properties with the corresponding properties of the same protein isolated from human and guinea pig milk. Immunochemical
quantitations showed that lactoferrin occurs in human neutrophilic leukocytes at the concentration of 3 µg per 106 cells. Tissue cultures from guinea pig bone marrow and spleen actively synthesized the protein, as shown both by net production
of lactoferrin and incorporation of labeled amino acids into the protein. Immunohistochemical data indicate that lactoferrin
first appears in myeloid cells at the stage of the promyelocyte.
ETHOLOGISTS developed the idea that many animal species evolved mechanisms to control aggression among their members. This control is by the use of signals to reduce the probability of attacks by the opponent before injurious or lethal combats occur1-3. Although there are observations that are best explained by the theory, no direct experimental evidence has been presented. We report here one type of experimental evidence that seems to fit the theory.
This chapter reviews the extensive literature on various transfer factors involved in cellular immunity. This is a very enlightening review of an area of immunology from which much is expected to apply in the future. The methodology is partially worked out for obtaining transfer factor, as well as some of the other materials, in sufficient purity for chemical analysis, and further results in this area are awaited with great interest. Transfer factor began with the demonstration that cellular transfer of tuberculin hypersensitivity can be accomplished in humans using viable leukocytes obtained from sensitive donors. These observations led to the finding that leukocyte extracts are as effective as viable cells in the transfer of delayed hypersensitivity in humans that allowed the search for the identity of the material(s) responsible for transfer factor activity to begin. The term “transfer factor” is used in the chapter to designate the specific factor or factors in leukocytes responsible for the transfer of delayed hypersensitivity. Transfer factor is also used to describe either the active moiety residing in specifically sensitive, viable blood leukocytes or is applied to the equivalent activity liberated from living cells mechanically ( lysis, freeze-thawing ) or immunologically (interaction with specific antigen), as well as to the more purified low-molecular-weight preparation separated from subcellular constituents by dialysis and concentrated by lyophilization.
Sheep red blood cells can surround, in vitro, some human peripheral blood lymphocytes in a formation called a rosette. The number of rosetteforming cells (RFC) in 50 normal persons had a wide range (4-40%). The organs of 13 human fetuses (11-19 wk conceptional age) were examined for the presence of RFC. The thymus possessed the highest percentage of RFC, the maximum being 65% of total thymocytes in two 15-16 wk fetal specimens. Blood RFC were always present and their number slightly increased in the oldest fetuses. The bone-marrow showed 0-8% in the six fetuses studied. RFC were found in the spleen around the 13th wk and in the liver around the 17th wk of gestation. These observations lead to the hypothesis that human blood RFC may be chiefly thymic derived. Studies of patients with immunological disorders support this hypothesis: one patient with Nezelof syndrome had no blood RFC and four patients with Wiskott-Aldrich syndrome had a low number of blood RFC (1 and 1.5%). Patients with acquired hypogammaglobulinemia showed a normal percentage of RFC. With the fetal thymocytes, the percentage of inhibition with anti-mu serum increased with the fetal age to become complete in the oldest fetuses studied. Incubation of the oldest fetal thymocytes or the blood lymphocytes with anti-gamma serum of anti-mu serum completely inhibited the rosette formation. These results suggest that mu-chain determinants are present on human fetal thymocytes and blood RFC. The significance of the presence of gamma-chain determinants on these cells is unclear.
Leucocytes obtained from a donor sensitive to Candida antigen contain a factor (transfer factor) which transfers this sensitivity to non-sensitive recipients. This material is present in the dialysate of leucocyte lysates and can be purified therefrom by gel filtration. This immunologically active component of transfer factor seems to be more potent than the crude dialysate, and contains a moiety which is readily labelled by culturing the leucocytes in the presence of 35S-labelled sodium sulphate. This labelled moiety has the properties of a short polypeptide chain joined to three or four R.N.A. bases.
Extract: The morphology and certain functions of human colostral cells were studied in vitro. Colostrum from 60 human females contained neutrophils, small lymphocytes, macrophages and occasional epithelial cells. The median concentration of neutrophils in colostrum from breast-feeding mothers was 150/mm3 as compared to 7000/mm3 in colostrum from those who did not breast feed.
PIP
Field observations are reported which support an assumption that human breast milk acts benefically through induced host resistance to infection. Though the mechanisms of this induced host resistance to infection are not clearly understood, this article suggests the following factors as important: specific antibodies to infectious agents, influences stimulating or inhibiting certain intestinal microorganisms, and nonspecific antimicrobial factors. Immunoglobulins are present in human milk, with IgA representing the most abundant; the role of complement and immunoglobulin in induction of resistance to infection is well-known. Since all immunoglobulins have antibody activity, it is conceivable that immunoglobulins provide the material for antibodies to various etiological agents; for example, serum IgA contains all types of antibodies; serum IgG has a wide variety of antibodies to viruses, rickettsiae, protozoa, H antigens of Salmonella, and bacterial antitoxins and incomplete Rh antibodies; and the IgM fraction contains antibodies to O antigens of Enterobacteriacae, Rh agglutinins, and syphilis reagins. Another aspect of human milk as an inducer of host resistance is the bifidus factor which promotes development of characteristic microflora; since the bifidobacteria metabolize a variety of sugars, producing large amounts of acetic and lactic acids and trace amounts of formic and succinic acids, these organisms affect pH and, hence, certain resistance factors. Other factors present in milk include lysozyme (muramidase), complement, interferon, and immune cells, all of which promote host resistance to infection.
The study concerned the binding of sheep red blood cells by human peripheral blood lymphocytes. The results indicate that the phenomenon (rosette formation) is probably dependent on thymus-derived lymphocytes (T lymphocytes) and not due to bone marrow lymphocytes (B lymphocytes). These data were obtained by a gradient centrifugation technique separating B lymphocytes from rosette-forming cells, by direct study of B lymphocytes and rosette-forming cells in the same preparations, and from the study of patients with selective immunodeficiency diseases and chronic lymphocytic leukaemia. Furthermore, inhibition of rosette formation was absent or very weak after incubation of lymphocytes with anti-immunoglobulin antisera, but considerable following their incubation with unspecific mitogens (phytohaemagglutinin, pokeweed mitogen, concanavalin A, anti-lymphocyte antiserum) and also after trypsin treatment. Incubation temperature was important for the frequency of rosettes, and experiments with metabolic inhibitors showed that to form rosettes the lymphocytes required an intact energy production.
TUMOR, rubor, calor et dolor, cum functione laesa: since the times of the ancients these have been the cardinal signs of inflammation, diagnostic of a reaction by the organism that was detrimental to its well-being. In 1883, Rudolph Virchow, the founder of cellular pathology, advised Metchnikoff to proceed with his researches cautiously, since "most pathologists do not believe in the protective role of inflammation."1 Metchnikoff proceeded, although not with caution, and his life's work provided the basis for the recognition that cellular as well as humoral mechanisms participate in immunity and immunopathology. There is now substantial evidence that cellular . . .
Peripheral blood lymphocytes from normal subjects as well as patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and active tuberculosis were studied for the relative distribution of bone marrow-derived lymphocytes (B-cells) and thymic-derived T-cells. B-cells were identified by direct immunofluorescence of surface Ig markers; T-cells were studied using rabbit antisera to pooled human fetal thymocytes absorbed with chronic lymphatic leukemia lymphocytes as a source of B-cells. In normal subjects, the sum of percentages of peripheral blood lymphocytes staining for surface Ig (B-cells) plus the percentage of cells staining with the absorbed antithymocyte antiserum closely approximated 100%. The mean value for percent B-cells among 51 normals tested was 22.9%+/-7.1; mean T-cells value was 75.3+/-13.95%. T-cell-specific antiserum stained 18% of normal human bone marrow lymphocytes, 42.5% of lymphocytes from normal spleens, and 98% of cells obtained from thoracic duct drainage of patients with RA. Specificity of antihuman thymocyte antiserum appeared to depend on the use of living cells. When patients with RA were examined, a wide range (14-98%) of peripheral blood T-cell values was found. Values for low percentages of peripheral blood T-cells appeared to correlate to some extent with severe clinical disease. In 11 of 36 RA patients, the sum of identifiable B- plus T-cells accounted for only 34-55% of peripheral blood lymphocytes. The identity of the remaining "null" cells could not be identified.3 of 24 SLE patients studied showed low percentages of peripheral blood T-cells, but no correlation could be drawn between T- to B-cell ratios and clinical disease activity. Among 21 patients with active tuberculosis, one had a low value for identifiable T-cells. No significant differences from normals in range or proportion of B-cells was identified in patients with active tuberculous infection.
Peripheral blood lymphocytes from 23 patients with active systemic lupus erythematosus (SLE) were serially studied. Changes in bone marrow-derived lymphocytes (B cells), as measured by surface Ig receptors and C3 receptors, and in thymus-derived cells (T cells) measured by rabbit T-cell-specific antiserum and E-binding techniques, were correlated with fluctuations in clinical disease activity and treatment. In normal controls B- and T-cell percentages remained relatively stable, although the situation in SLE was much more labile. A relative and absolute decrease in T lymphocytes and cells bearing a receptor for C3 was found in active lupus. Absolute numbers of cells bearing surface Ig were decreased to a lesser extent, whereas the proportion of these cells was increased. It is postulated that the increase in autoantibody formation and diminished delayed hypersensitivity seen in systemic lupus may be due to a loss of T-lymphocyte function.
Further evidence is given of a mitogenic response to phytohemagglutinin by human B lymphocytes. Immunofluorescent staining of unfractionated lymphocytes, and thymidine uptake by column purified populations of B and T cells, indicates that the dose time response of these two populations is comparable. Phytohemagglutinin transformation of B lymphocytes appears to be reduced or absent in chronic lymphocytic leukaemia.
Human chorionic gonadotropin completely inhibits the response of lymphocytes to phytohemagglutinin. The effect is both reversible and noncytotoxic. These observations support the theory that the fetus is accepted because human chorionic gonadotropin represents trophoblastic surface antigen and blocks the action of maternal lymphocytes.
Lymphocytes from pregnant women and non-pregnant individuals were cultured under the stimulus of phytohaemagglutinin in the presence of their own and heterologous (allogeneic) sera. The results indicate that heterologous sera have an inhibitory effect on the lymphocyte transformation rate and suggest that the inhibitory property is more powerful in pregnant and fetal sera. Conversely, the addition of heterologous non-pregnant sera to cultures of pregnant lymphocytes increases their transformation rate. These findings suggest that there is a serum inhibitor in pregnancy and this finding may be relevant to the survival of the fetal allograft.
The distribution of peripheral blood lymphocytes that contain surface Ig has been studied by means of immunofluorescence in humans. Normal individuals, individuals with sex-linked and acquired agammaglobulinemia, selective IgA deficiency, cellular immune deficiencies, and individuals with chronic lymphatic leukemia (CLL) were studied. Approximately 28% of the peripheral blood lymphocytes from normal individuals contained surface Ig. On an average 15% contained IgG, 6%, IgA, and 8%, IgM; and the kappa: lambda ratio was 2:1. Lymphocytes from patients with CLL appeared to be "monoclonal" in that the cells from a given individual had a single Ig associated with them (e.g., kappa IgM). In three-quarters of the cases the H chain class was IgM; in the remaining one-quarter no H chain could be detected on the cell surface. The L chain class was kappa in 12 cases and lambda in 8. Four patients with sex-linked agammaglobulinemia and one with "acquired" agammaglobulinemia had markedly decreased numbers of cells with surface Ig (0-4%). In contrast, the three patients with selective IgA deficiency and no detectable serum IgA contained normal numbers of cells (6-8%) with surface IgA. Five patients with cellular deficiency states, including two with Wiskott-Aldrich syndrome, contained a normal or low percentage of cells with surface Ig.
Extract: The production of IgG, IgA, IgM, β1c, and the secretory piece (SP) by the cells of human colostrum was investigated by radioimmunoelectrophoresis. The synthesis of IgA and β1c by the cells was demonstrated. No evidence, however, for the formation of IgG of IgM by colostral cells was found. Similarly, no radiolabeling was found over the free SP immunoprecipitin lines, but labeling was detected over the bound SP immunoprecipitin lines. This many have been due to covalent binding of the SP with radiolabeled α chains of secretory IgA.