Regional immunology (Reg Immunol)

Publications in this journal

• Article: The kinetics of migration of murine CD4 and CD8 lymphocytes in vivo.

  L L Fisher, C A Ottaway
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  ABSTRACT: Lymphocytes from the mesenteric lymph nodes (MLN) of mice were enriched for CD4+ and CD8+ T cell populations, labeled with (51Cr) sodium chromate, and transferred to the bloodstream of syngeneic recipients. The time course of migration of the labeled cells from the blood to the secondary lymphoid organs of the recipients was investigated. CD4+ and CD8+ subpopulations differed in their profile of appearance in different lymphoid organs. Computer assisted analysis of the observations was used to obtain quantitative estimates of the rate of clearance of the lymphocytes from the blood into the tissues, and the rate of departure of the cells from the tissues. In the spleen, CD4+ lymphocytes were cleared from the blood about one and one-half times faster than CD8+ lymphocytes, but the CD8+ cells were retained longer. Inguinal nodes (IN), MLN, and Peyer's patches (PP) showed a consistent ability to clear CD4+ cells from the blood at a rate approximately 2.5 x greater than that for CD8+ cells, but the retention of the lymphocytes in these tissues varied with lymphocyte phenotype and the organ concerned. CD4+ lymphocytes were retained longer in PP and MLN than in IN, whereas CD8+ cells were retained longer in IN and MLN nodes than in PP. We conclude that the rate of clearance of lymphocytes into secondary lymphoid organs from the blood varies in a regular way with T cell phenotype and that organ specific sorting of T subpopulations also proceeds after the cells are admitted to the tissues.
  Regional immunology 3(3):156-62.
• Article: The mucosal immune system: features of inductive and effector sites to consider in mucosal immunization and vaccine development.

  H Kiyono, J Bienenstock, J R McGhee, P B Ernst
  Regional immunology 4(2):54-62.
• Article: Interferon-gamma, Staphylococcus aureus, and lipopolysaccharide/silica enhance interleukin-1 beta production by human corneal cells.

  N B Shams, M M Sigel, R M Davis
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  ABSTRACT: Cultures derived from human corneo-scleral rims remaining after a central corneal button had been removed for transplantation, revealed two types of cells on light microscopy: One with typical epithelial morphology and the other resembling fibroblasts. Both cell types contained keratin filaments in early passage and were therefore considered epithelial in nature. The fibroblast-like cells were designated fibroblast-like epithelial cells (FLE) while the typical epithelial cells were referred to as E-type. Both E and FLE cells constitutively produced an IL-1-like factor as determined by thymocyte proliferation assay and IL-2 induction in EL-4 lymphoma cells. Moreover, the supernatants from these cells potentiated concanavalin A (Con A)-primed mitochondrial oxidative metabolism in thymocytes, as indicated by the tetrazolium salt reduction assay (MTT) and this effect was significantly neutralized with monoclonal anti-IL-1 beta. The release of biologically active IL-1 beta by the FLE cells is another characteristic (in addition to the presence of keratin) distinguishing these cells from fibroblasts which do not release biologically active IL-1 beta. Using an ELISA, specific for IL-1 beta, there was clear cut evidence for increased production of this cytokine by FLE cells in response to human recombinant gamma-interferon (IFN-gamma), Staphylococcus aureus, and lipopolysaccharide (LPS) in combination with silica (LPS/silica). Time studies with IFN-gamma and LPS/silica demonstrated that enhanced production was time dependent and that IL-1 beta was primarily cell associated. The results indicate that human corneal E- and FLE-type cells can produce and release IL-1 and that FLE cells can be induced by inflammatory mediators to increase production of IL-1 beta.
  Regional immunology 2(3):136-48.
• Article: Characterization of human IgG subclasses within intraocular compartments.

  J C Waldrep, J R Schulte
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  ABSTRACT: Multiple factors regulate the distribution of the human immunoglobulin (Ig) G1-4 subclasses within different compartments of the human eye. An experimental protocol was developed for analysis of IgG1-4 using a combination of high pressure liquid chromatography (HPLC) chromatofocusing and an enzyme-linked immunosorbent assay (ELISA). IgG in the human cornea, aqueous humor, and vitreous was extracted and fractionated by chromatofocusing in a pH 9.0-6.0 gradient. A monoclonal antibody based capture-type ELISA was used to determine the IgG1-4 subclass distribution within the fractions. Histograms generated were then used to determine the IgG1-4 subclass profile for each intraocular compartment. Results indicate that the IgG species are similar but unique for each compartment. In contrast to the distribution for normal serum/plasma IgG1-4, the intraocular profiles were extremely heterogeneous and variable. These results indicate that the intraocular compartments are in a state of disequilibrium. There are apparently both common and unique factors that regulate the distribution of individual IgG subclass species for each intraocular compartment. In addition to differences in fluid dynamics, tissue biochemistry, and barrier properties, parameters, unique to the IgG subclass species may also profoundly effect intraocular localization and retention.
  Regional immunology 2(1):22-32.
• Article: Phagocytosis of particulate antigens by corneal epithelial cells stimulates interleukin-1 secretion and migration of Langerhans cells into the central cornea.

  J Y Niederkorn, J S Peeler, J Mellon
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  ABSTRACT: Under normal physiological conditions, the central corneal epithelium is devoid of Ia+ Langerhans cells. However, a variety of stimuli can induce the migration of peripheral Langerhans cells into the central regions of the cornea. In the present study, Langerhans cell migration was induced by the instillation of either sterile latex beads or formalin-killed Staphylococcus aureus into shallow incisions in the central corneal epithelium. Langerhans cells could be detected in the central cornea as early as 24 hours following instillation of either latex beads or S. aureus and remained for at least 6 weeks. Phagocytosis of latex beads by corneal epithelial cells was demonstrated in vivo and in vitro. Moreover, phagocytosis of latex beads stimulated corneal epithelial cells to secrete increased amounts of interleukin-1 (21-83% increase). The centripetal migration of peripheral corneal Langerhans cells in response to phagocytosis of latex beads could be mimicked by injecting as little as 0.001 units of human purified interleukin-1 (IL-1) into the central corneal epithelium. The IL-1 induced chemotaxis could be blocked by coinjection of anti-IL-1 antibody but not irrelevant antibody. The findings indicate that the exclusion of Langerhans cells from the central corneal epithelium is a dynamic process that can be regulated by the resident corneal epithelial cells themselves and raises the possibility that corneal epithelial cells and Langerhans cells collaborate in antigen processing within this organ.
  Regional immunology 2(2):83-90.
• Article: Intraduodenal application of cholera holotoxin increases the potential of clones from Peyer's patch B cells of relevant and unrelated specificities to secrete IgG and IgA.

  D A Lebman, J A Fuhrman, J J Cebra
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  ABSTRACT: Although it has been established that cholera toxin is both an effective mucosal immunogen and adjuvant, the mechanism by which it acts has not been determined. The relative contributions of the pharmocologic and binding capacities of holotoxin have been assessed by comparing holotoxin, acid dissociated B subunit, and B subunit from the Texas Star variant, which is deficient in production of the A subunit, for their ability to generate toxin-specific precursors in vivo that give rise to clones secreting exclusively IgA in vitro. In this way we demonstrated that although B subunit is as immunogenic as the holotoxin, pharmacologic activity appears to play a role in the generation of IgA-committed precursors. In addition, intraduodenal (i.d.) application of holotoxin can act to alter the isotype display of previously primed B cells in Peyer's patches (PP) specific for a chemically unrelated hapten resulting in an increase in the proportion of their clones that secrete IgA or IgG and a decrease in the proportion that secrete IgM following antigen-dependent in vitro clonal expansion. Intraduodenal application of cholera toxin did not appear to nonspecifically increase the proportion of IgA-committed precursors as judged by staining for sIgA and mRNA alpha levels. However, following i.d. application of cholera toxin, an overall downward shift in the levels of sIgD and s kappa was observed in the PP B cell population. We suggest that the holotoxin can nonspecifically affect the isotype display of PP B cells by altering their responsiveness to the stimuli present in the in vitro cultures.
  Regional immunology 1(1):32-40.
• Article: Locally and systemically derived natural killer cells participate in defense against intranasally inoculated influenza virus.

  J Stein-Streilein, J Guffee, W Fan
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  ABSTRACT: Previous studies have indicated that the pulmonary natural killer (NK) cells are important in early defense against influenza virus (PR8/34) infection by the intratracheal route. Since the natural route of virus entry into the lung is via the upper respiratory tract, the present study was to elucidate the role of NK cells in early defense to intranasal (IN) inoculation of influenza virus. Rabbit anti-Asialo GM1 (RAGM1) serum was administered by IN, intravenous (IV), or both routes, 24 hours before IN infection with a median lethal dose (LD50) of PR8/34 previously shown to kill 50% of the inoculated mice (B6D2F1 or nu/nu) by day 8. IN or IV inoculation of 20 microliters of RAGM1 (whole rabbit serum) had no effect on the survival of the mice to virus administered by the IN route. When RAGM1 was given IV (10 microliters) and IN (10 microliters) 24 hours before PR8/34 IN, 60% of the mice (B6D2F1 or nu/nu) died by day 4. Influenza virus titers were at least one log higher in the lungs of B6D2F1 mice and two logs higher in IN/IV NK-depleted nu/nu mice than in lungs of mice who received RAGM1 by IV or IN route alone or who were untreated. NK activity was depleted in lungs but not blood of mice treated IN with RAGM1 and was depleted in both lungs and blood if RAGM1 was given IV. These data support the hypothesis that NK cells are important to early defenses against influenza virus in the upper respiratory tract and that they are derived from both a local and systemic population of NK cells.
  Regional immunology 1(2):100-5.
• Article: Immunoregulatory functions of BCG-elicited alveolar macrophages in cell-mediated immune responses.

  J U Igietseme, H B Herscowitz
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  ABSTRACT: The pulmonary alveolar macrophage (AM) has been shown to play an important role in maintenance of the lung. However, the function of AM in specific cell-mediated immune responses has not been studied in detail. By employing both functional and immunologic assays, the present studies were undertaken to investigate the accessory and immunoregulatory functions of BCG-elicited AM in cell-mediated immune responses in vitro. AM, at concentrations of from 1.0 to 10%, could replace the accessory function of splenic adherent cells in the biological and functional activation of purified T-lymphocyte populations stimulated by mitogen and allogeneic cells as well as for the generation of cytotoxic effector cells. In unfractionated lymphoid cell cultures, AM were found to exert pronounced dose-dependent suppressive effects on mitogen and allogeneic cell stimulation. AM-mediated suppression of cell-mediated immune responses required metabolically active AM, but it did not require direct physical contact between AM and the stimulated lymphocytes. Furthermore, kinetic studies suggested that AM-mediated suppression was a time-dependent event that affected the afferent phase of T-cell activation by arresting the development of a primary activation signal. Moreover, the biological manifestation of suppression was characterized by inhibition of blast transformation, a decline in interleukin-2 receptor expression, as well as decreased proliferation and differentiation of precursors into effector cells (cytotoxic T-lymphocytes). These results suggest that BCG-elicited AM are capable of both accessory and regulatory functions in cell-mediated immunity.
  Regional immunology 1(3):172-81.
• Article: Cellular analysis of functional mononuclear cells from chronically inflamed gingival tissue.

  M L McGhee, T Ogawa, A M Pitts, Z Moldoveanu, J Mestecky, J R McGhee, H Kiyono
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  ABSTRACT: Functional mononuclear cells from chronically inflamed gingiva of different stages of adult periodontitis (AP) were isolated by using a novel enzymatic dissociation method and extensively characterized at the single cell level. Fresh tissues obtained following surgery were cut into 1-2 mm3 pieces, washed free of residual blood cells and dissociated with the neutral protease enzyme Dispase. Mononuclear cells were then obtained by separation on a mini-Ficoll-Hypaque gradient. This procedure yields an average of 2 x 10(6) cells/400 mg of tissue. The viability of unfractionated cells immediately after enzymatic dissociation was greater than 94%, and after Ficoll-Hypaque gradient separation this was greater than 99%. The relative percentages of lymphocytes, plasma cells, monocytes (MN)/macrophages (M phi) and granulocytes were determined on cytospin preparations by Wright's-Giemsa staining. Cell differential analysis showed that lymphocytes and MN/M phi predominated at all stages of disease, while the frequency of plasma cells increased with the severity of disease. Numbers of plasma cells specific for IgG, IgA, or IgM were determined by immunofluorescence using fluoresceinated anti-heavy chain specific antibodies. The majority of Ig-containing cells were of the IgG isotype, followed by significant numbers of IgA and few IgM-positive cells. IgG, IgA, and IgM secreting cells were also determined by the ELISPOT assay. Total numbers of spot forming cells (SFC) were measured in both the moderate and advanced stages of disease, and the major SFC isotype was IgG followed by IgA; essentially no IgM SFC were detected. Furthermore, higher numbers of IgG and IgA SFC were noted in the advanced stage when compared with the moderate stage of disease.(ABSTRACT TRUNCATED AT 250 WORDS)
  Regional immunology 2(2):103-10.
• Article: Comparison of systemic and local immune responses after multiple pulmonary antigen exposures.

  M J Mason, N A Gillett, D E Bice
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  ABSTRACT: Both systemic and local immune responses were evaluated in five male cynomolgus monkeys after a primary and multiple secondary challenges of the lung with sheep red blood cell (SRBC) antigen. Comparisons of peripheral blood and pulmonary antibody-forming cell (AFC) responses were made after primary and multiple secondary intrapulmonary challenges with SRBC. The primary immune response to SRBC was characterized by a low lung to blood AFC ratio. This ratio, however, increased successively with each challenge, resulting in a 15-fold increase after the third exposure of the lung to SRBC. These data suggest a decreasing contribution of systemic AFC to the lung response with each successive challenge. Histopathologic and autoradiographic evaluation was done to compare a newly immunized lobe, a multiply challenged lobe, and a saline instilled lobe. Large localized perivascular lymphoid follicles appeared in the lung lobe challenged multiple times. About 17% of these large mononuclear cells had taken up 3H-thymidine compared to 5% in the newly immunized lobe and 3.8% in the control lobe. The number of proliferating cells appeared to reflect the degree of immune responsiveness within each lung lobe. Together these data support a hypothesis that after the primary immune response is established, the secondary immune response is generated by local clonal expansion of antigen-specific lymphocytes within the lung.
  Regional immunology 2(3):149-57.
• Article: Effects of CD4+ and CD8+ T-lymphocyte depletion on the induction and expression of herpes simplex stromal keratitis.

  C K Newell, D Sendele, B T Rouse
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  ABSTRACT: The immunopathological role of T-lymphocytes in Herpes Simplex Virus type-1 (HSV-1) induced stromal keratitis (SK) has been well established, however, many questions still remain as to the actual mechanism(s) involved in the expression of this disease. To address this issue we have depleted mice of CD4+ and CD8+ T-lymphocytes at various stages of the disease and evaluated the effect of this therapy on the clinical outcome of the disease. We found that depleting mice of CD4+ T-lymphocytes either ameliorated the disease or halted its further progression. In contrast, depleting animals of CD8+ T-lymphocytes exacerbated or had no effect on the outcome of the disease. The results of this study suggest that CD4+ T-lymphocytes are involved in both the induction and the expression of SK, and CD8+ T-lymphocytes may serve some regulatory function in this disease.
  Regional immunology 2(6):366-9.
• Article: Influence of microenvironmental factors on human Langerhans cell function in vitro.

  A Demidem, J R Taylor, S F Grammer, J W Streilein
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  ABSTRACT: When murine epidermal cells are placed in tissue culture for 48-72 hr, the Langerhans cell subpopulation acquires a markedly enhanced capacity to activate autologous as well as allogeneic T cells. This change is mediated largely, if not exclusively, by GM-CSF derived from keratinocytes present in the same cultures. When human epidermal cells are cultured under similar conditions, Langerhans cells display much more limited functional changes. We have investigated whether the relative failure of human Langerhans cells to change their functional program in vitro is due to (a) production of prostaglandins by keratinocytes during functional assays (which would inhibit T cell proliferation), and/or (b) relative deficiency of GM-CSF in the 72-hr epidermal cell culture. Freshly procured and cultured (72 hr) human epidermal cells were used as stimulators for allogeneic T cells in cultures to which indomethacin was added to block prostaglandin production. Under these circumstances, the allostimulatory capacity of cultured cells was enhanced, although there was no significant increase in the stimulatory properties of fresh cells, implying that prostaglandins may account in part for the relatively poor antigen presenting properties of cultured human cells. In addition, human epidermal cells were cultured with or without exogenous GM-CSF prior to being used as stimulators for autologous and allogeneic T cells. Following exposure to GM-CSF, cultured human epidermal cells acquired markedly enhanced T cell-activating properties, rivaling those reported for cultured murine epidermal cells.(ABSTRACT TRUNCATED AT 250 WORDS)
  Regional immunology 3(6):297-304.
• Article: Inhibition of DNA synthesis in human peripheral and lamina propria lymphocytes by 5-aminosalicylic acid and hydrocortisone.

  Y Elitsur, C P Freedland, G D Luk
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  ABSTRACT: 5-Aminosalicylic acid (5-ASA) and hydrocortisone (HC) have potent antiinflammatory activity, and are commonly used in the treatment of patients with inflammatory bowel disease. Both agents have been reported to modulate immune function in human and animal models. We investigated the effect of 5-ASA and HC on cell proliferation of normal human peripheral blood lymphocytes (PBL) and normal colonic lamina propria lymphocytes (LPL). Both 5-ASA (0.01-4 mM) and HC (0.01-10 microM) significantly inhibited 3H-thymidine incorporation (Thd) into PBL and LPL in a dose dependent manner. Maximal inhibition occurred with the highest concentration tested of both 5-ASA and HC. We found a significantly greater inhibition of DNA synthesis for 5-ASA (4 mM) upon LPL vs. PBL (94 vs. 32%, p less than 0.001). However, DNA synthesis inhibition by HC was similar in PBL and LPL. The profoundly greater inhibitory effect of 5-ASA upon LPL suggests a specific modulation of the mucosal immune system.
  Regional immunology 3(1):56-61.
• Article: Sodium periodate treatment modulates the accessory and regulatory functions of alveolar macrophages in T-cell responses.

  F T Duque, H B Herscowitz
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  ABSTRACT: It is known that murine alveolar macrophages function inefficiently as antigen-presenting cells for the in vitro activation of macrophage-depleted T-lymphocyte populations and that they suppress antigen-stimulated lymphoproliferative responses in a dose-dependent manner. The present studies were carried out to determine whether oxidation of the alveolar macrophage surface could alter these activities. Viable alveolar macrophages were treated with varying concentrations of sodium periodate then cultured with either unfractionated or adherent cell-depleted lymphocyte populations obtained from BCG-immunized animals and challenged with PPD. It was demonstrated that moderate oxidation of the alveolar macrophage surface with sodium periodate abrogated their ability to suppress antigen-stimulated proliferation of unfractionated lymphocyte populations, as well as resulted in initiation of lymphoproliferation in an antigen-stimulated, adherent cell-depleted spleen cell populations. It is conceivable that mild oxidation of the alveolar macrophage membrane results in a more favorable interaction between the alveolar macrophage and responding T-lymphocytes, thereby reversing the alveolar macrophage-mediated suppression of antigen-stimulated lymphoproliferation and allowing for effective antigen-presenting function of these cells.
  Regional immunology 2(3):129-35.
• Article: Influences of the microenvironment on B-cell responses of wasted mice: comparison of Peyer's patches and mesenteric lymph nodes.

  G E Woloschak, C J Krco, M Rodriguez
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  ABSTRACT: Wasted mice bear an autosomal recessive mutation (wst/wst) that gives rise to neurologic abnormalities and immunologic deficiency evident as early as 21 days of age. Features of the immune deficiency include absence of plasma cells at mucosal sites, reduction in weight of thymus and spleen relative to body weight, and low serum levels of some immunoglobulin (Ig) isotypes. In these experiments, we have examined parameters of B-cell function in Peyer's patches (PP) and mesenteric lymph nodes (MLN) of wst/wst and age-matched control mice. Our results established the following abnormalities in PP but not MLN of wst/wst mice relative to controls: (1) reduced mitogenic responses to lipopolysaccharide, (2) decreased percentages of Ig+ cells, (3) increased percentages of B-cells bearing large amounts of surface Ig ("very bright" Ig+ cells), and (4) reduced levels of all Ig-specific messenger ribonucleic acids (mRNAs) except alpha-mRNA. The only immunologic abnormality consistently expressed in lymphocytes from both MLN and PP of wst/wst mice is reduced levels of IgA+ B cells and alpha-mRNA. Morphologic studies revealed no consistent abnormalities in lymph nodes, spleens, livers, or PP of wst/wst mice when compared to littermate controls. These results support the idea that some of the immunologic abnormalities reported in wst/wst mice are due to microenvironmental influences. Of the parameters examined in this report, only reduced IgA (mRNA levels and percentage of positive cells) is consistently observed in PP, MLN, and spleen.
  Regional immunology 1(3):163-71.
• Article: Regulation of mucosal immune responses by T lymphocytes: the effect of chronic CD4+ T cell deficiency on IgA synthesis.

  J Mega, K Fujihashi, H Kiyono
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  ABSTRACT: Mechanisms of immune defence at the mucosal surface has been elucidated by recent advances in molecular and cellular immunology. IgA is undoubtedly the most important defense factor in the mucosal immune system. It has been shown that T cells are essential for the induction and regulation of IgA synthesis. In T cell regulation of IgA synthesis, various cytokines (e.g., TGF-beta, IL-2, IL-5, and IL-6) which are secreted by CD4+ T cells, play important roles for the induction and regulation of IgA isotype switching and terminal differentiation of sIgA+ B cells to become IgA producing cells. The chronic treatment of mice with anti-CD4 mAb induced a market deficiency of CD4+ T cells in both mucosal and systemic tissues. IgA plasma cells were significantly reduced in treated mice when compared with normal mice (greater than 80% reduction), while the numbers of sIgA+ B cells in IgA inductive sites (e.g., PP) remained normal. CD4+ Th cells are a critical element for the induction of appropriate IgA responses in mucosal associated tissues. Elucidation of the precise cellular and molecular network for the regulation of mucosal immune defense system is important and useful for the consideration of prevention of infectious diseases. In this regard, the effective and sophisticated mucosal administration of vaccines using the concept of the common mucosal immune system should induce effective immune responses which prevent the pathogen from entering the host through large surface areas of mucosal membranes. This goal cannot be achieved without a more complete understanding of regulatory T cells and cytokines for mucosal immune responses.
  Regional immunology 4(2):70-8.
• Article: Immune regulation of metastasis from in vivo derived syngeneic murine melanoma.

  R Harning, G C Koo, J Szalay
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  ABSTRACT: In earlier work, we demonstrated that in vivo derived B16F10 tumor cells metastasize aggressively from intracameral (ic) and subcutanous (sc) sites, colonizing the lungs and lymph nodes. Natural killer (NK) cells play an important role in metastasis from ocular tumors. Treatment of mice with MoAb PK136, a highly specific anti-NK antibody, altered the pattern of metastasis; metastases appeared in the lungs, adrenal glands, liver, and spleen. Treatment with cyclophosphamide (Cy) did not affect survival or metastasis, but combined treatment with the immunomodulator Linomide (LS2616) and Cy decreased metastasis and increased survival. In the present study, we examine the role played by NK cells in regulating metastasis of sc tumors. Treatment of mice with LS2616 enhanced NK cell activity and decreased metastasis. Treatment with MoAb PK136 decreased survival and increased metastasis, but did not affect the pattern of metastasis. Treatment with 25 mg/kg Cy alone resulted in a decrease in growth of the primary tumor, increased survival, and decreased metastasis. Combined treatment with LS2616 and Cy was very effective in decreasing metastasis, increasing survival, and affecting cures (30%). In summary, our experiments demonstrate the importance of NK cells in regulating metastasis from sc tumors of in vivo derived B16F10 melanoma and demonstrate the effectiveness of LS2616 and low doses of Cy on metastasis and survival. In combination with earlier work, the present experiments demonstrate: 1) that modulation of NK activity differentially affects metastasis from sc and ic compartments, and 2) that regional differences in the location of the primary tumor may determine the effectiveness of treatment with Cy.
  Regional immunology 3(2):97-102.
• Article: Induction of Fc alpha receptor expression on T cells from murine Peyer's patch, spleen and thymus.

  D Char, W K Aicher, J R McGhee, J H Eldridge, K W Beagley, H Kiyono
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  ABSTRACT: It has been shown that Fc alpha receptor-positive (Fc alpha R) lymphocytes function in the regulation of IgA responses. In the present study, we have compared the frequency of Fc alpha R+ T cells in IgA inductive sites, e.g., the Peyer's patches (PP) with a systemic tissue, the spleen (SP), and the progenitor tissue, thymus. CD3+, Thy-1+ T cells were incubated with purified IgA and then counterstained with fluorescence conjugated anti-mouse alpha-chain specific antibody. Flow cytometry (FACS) analysis showed a distinct subset of Fc alpha R+ T cells in both PP (5-10%) and SP (1-4%), while essentially no Fc alpha R+ T cells were found among isolated thymocytes. The Fc alpha R+ T cells in PP were associated with the population of large, blast cells as determined by forward angle light scatter analysis. The expression of the Fc alpha R could be induced or augmented by activation of T cells with anti-CD3 antibody. Incubation of thymocytes with anti-CD3 antibody resulted in the induction of thymocyte blasts and in Fc alpha R expression in this large cell population. Treatment of splenocytes with anti-CD3 also induced T cell blasts and enhanced the frequency of Fc alpha R+ T cells in this large cell population. Furthermore, when splenic T cells were separated into large blasts and small, resting T cell fractions on Percoll-gradients and then incubated with anti-CD3 antibody, the enhancement of Fc alpha R+ T cells was only noted in the latter cell population.(ABSTRACT TRUNCATED AT 250 WORDS)
  Regional immunology 3(5):228-35.
• Article: The use of the hu-PBL-SCID mouse model to study lymphocyte homing and responsiveness to recall antigens.

  C Lue, H Kiyono, K Fujihashi, J R McGhee, J Mestecky
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  ABSTRACT: SCID mice were injected intraperitoneally with human peripheral blood lymphocytes (PBL) that had been previously stimulated with pokeweed mitogen (PWM) for two days to generate activated B cells. After two weeks, serum and gut washes obtained from SCID mice reconstituted with human PBL (hu-PBL-SCID mice) contained human IgA, IgG, and IgM indicating the successful survival of human lymphoid cell grafts in both mucosal and nonmucosal tissues of the SCID mice. Human IgA plasma cells could be detected by immunofluorescence microscopy in the lamina propria of the small intestine, while IgM plasma cells predominated in the spleen. The results suggested that PWM-activated plasma cell precursors homed to the spleen and the lamina propria of the SCID mouse where they differentiated into plasma cells. In vivo stimulation of hu-PBL-SCID mice with diphtheria and tetanus toxoids (DT/TT) elicited a primary (IgM) immune response pattern rather than a secondary (IgG) response. Antigen-specific antibody-secreting cells were found in the spleen and lamina propria after immunization. The microenvironment of the hu-PBL-SCID mice may select virgin B cells subsets over memory B cell clones.
  Regional immunology 4(2):86-90.
• Article: Ontogeny of lymphocyte activating factors in conventional and germfree rats.

  T Granholm, B Fröysa, T Midtvedt, O Söder
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  ABSTRACT: Immune regulatory cytokines have previously been demonstrated outside the immune system in, e.g., the skin, testis, and brain, and suggested to serve as barrier site defence mediators. We have recently demonstrated constitutive production of an interleukin-1 (IL-1)-like factor(s) in the proximal gastrointestinal tract, liver, and placenta of adult rats. In the present study we have investigated the developmental production of IL-1-like factors in these tissues in conventional rats and in rats raised under germfree conditions. Slight IL-1 bioactivity was detected at a gestational age of 18 days in the skin and esophagus, and on day 19 significant amounts were detected in the skin, tongue, esophagus, and stomach. The activity was maximal at gestational day 20-21, and stationary at all later time-points. The placenta and liver contained high amounts of IL-1 activity throughout fetal life. The brain showed a transient appearance of IL-1 bioactivity at a gestational age of 19-21 days, but no activity at later time-points. Germfree rats showed no significant difference in the ontogeny or amounts of IL-1 bioactivity compared to conventional rats. It can be concluded that the IL-1 bioactivity appears at a definite time point in all measured tissues in fetal life and that exposure to microbial antigens does not seem to influence its production. As the investigated tissues show a high proliferation rate, it is tempting to suggest that the detected IL-1 (-like factors) might have an alternate function in these tissues, e.g., to serve as growth factors.
  Regional immunology 4(4):209-15.