Andrea Introini

Alfasigma

Background SARS-CoV-2 transmission mainly occurs through exposure of the upper airway mucosa to infected secretions such as saliva, which are excreted by an infected person. Thus, oral mucosal immunity plays a central role in the prevention of and early defense against SARS-CoV-2 infection. Although virus-specific antibody response has been extensi...

The association between the use of the injectable contraceptive depot medroxyprogesterone acetate and HIV-1 susceptibility has been addressed mainly in respect to the changes occurring in the female genital mucosa and blood. However, one of the main sites of HIV-1 pathogenesis is lymphoid organs. To investigate the immunoregulatory effect of medrox...

Discovering new medicines relies on the ability to identify and prioritize promising new drug targets. Unfortunately, biomedical science tends to focus on only a small fraction of the human genome and leaves large parts unexplored. This chapter highlights the importance of applying high‐quality chemical probes and chemogenomic libraries to evaluate...

Histocultures allow studying intercellular interactions within human tissues, and they can be employed to model host-pathogen interactions under controlled laboratory conditions. Ex vivo infection of human tissues with human immunodeficiency virus (HIV), among other viruses, has been successfully used to investigate early disease pathogenesis, as w...

Problem Susceptibility to HIV is associated with the menstrual cycle and vaginal microbiome, but their collective impact on vaginal inflammation remains unclear. Here, we characterized the cervicovaginal proteome, inflammation, and microbiome community structure and function during the menstrual cycle. Method of study Cervicovaginal secretions wer...

Gelfoam® histoculture provides a valuable tool for experimental studies of normal and pathological tissue physiology. It allows us to understand cell-cell interactions by mirroring their original spatial relationship within body tissues. Gelfoam® histoculture can be employed to model host-pathogen interactions mimicking in vivo conditions in vitro....

Background: Genital mucosa is the main portal of entry for various incoming pathogens, including HIV, hence an important site for host immune defenses. Tissue-resident memory T (TRM) cells defend tissue barriers against infections and are characterized by expression of CD103 and CD69. Here we describe the composition of CD8+ TRM cells in the ectoc...

[This corrects the article DOI: 10.1371/journal.ppat.1006402.].

The most immediate and evident effect of mucosal exposure to semen in vivo is a local release of proinflammatory mediators accompanied by an influx of leukocytes into the female genital mucosa (FGM). The implication of such response in HIV-1 transmission has never been addressed due to limitations of currently available experimental models. Using h...

Seminal cytokine carry-over in ectocervical tissue explant conditioned medium (ECM). Donor-matched ectocervical tissue explants were processed fresh or inactivated with paraformaldehyde (PFA), and incubated with culture medium (CM vs. CM-PFA) or seminal plasma (SP vs. SP-PFA) 50% for 12 h, followed by an additional 12 h-incubation with medium only....

Gating strategy used for flow cytometric analysis of peripheral blood leukocytes in transmigration assays. Acquired events were identified as: 1) counting beads; 2) leukocytes (CD45+); 3) granulocytes; 4) neutrophils (CD16+CD66b+); 5) T cells (CD16-CD3+); 6) classical monocytes (CD16-CD14+). (TIF)

Cell viability and tissue architecture of cultured ectocervical explants. Sections were obtained from donor-matched ectocervical tissue explants: A and D) incubated seminal plasma (SP) 50% for 12 h; B) incubated with culture medium (CM) for 12 h; C) snap-frozen immediately after dissection. A, B, and C) Sections were processed to reveal chromatin f...

Imaging of HIV-1 replication in ectocervical tissue explants. Representative section of a tissue explant infected with HIV-1BaL harvested on day 18 post-infection (top) and uninfected uncultured ectocervical tissue (bottom). In situ HIV RNA hybridization signal is shown in red. Scale bars = 200μm; 50μm (magnification). (TIF)

Primers used for quantitative real-time PCR. N/A not applicable. (PDF)

Effect of progesterone-based drug use on HIV-1BaL infection. Infection of ectocervical explants with HIV-1BaL was independently performed after an initial treatment with seminal plasma (post-SP, red), or in the presence of SP (SP-mix, blue) (see S1B Fig). Experiments were grouped based on progesterone-based drug use of tissue donors. N-fold change...

IL-6 and CXCL8 concentration in ectocervical tissue explant conditioned medium (ECM) of excluded experiments. A) Cytokine concentration (ng/ml) was measured in explant conditioned medium (ECM) of ectocervical explants incubated with culture medium (CM) or seminal plasma (SP) 50%, in the presence or absence of indomethacin (indo) 10μM for 4 or 12 h,...

Indomethacin titration. A) Cytokine concentration (ng/ml) was measured in explant conditioned medium (ECM) of ectocervical explants incubated with culture medium (CM, white), seminal plasma (SP, red) 50% or lipopolysaccharides (LPS, gray) 2μg/ml, in the presence or absence of indomethacin (indo) 10μM or 30μM, for 12 h, followed by an additional 12...

Study experimental design. A) Donor-matched explants of ectocervical mucosa were mounted in agarose medium with the epithelial side upwards, and incubated with seminal plasma (SP) or culture medium (CM) for 2, 4 and 12 h. Tissue integrity and cell viability were assessed by in situ TUNEL assay in explants harvested after 12 h-incubation with SP50%...

Expression levels of the chemokine receptors CXCR1 and CXCR2 on transmigrated granulocytes. Mononuclear cells and granulocytes were isolated from blood, pooled (i.e. peripheral blood leukocytes (PBL)) and incubated in a transwell system for 2 h with explant conditioned medium (ECM) from donor-matched ectocervical explants incubated with culture med...

Cytokine concentration in ectocervical tissue explant conditioned medium (ECM) and seminal plasma pools. Cytokine concentrations (pg/ml) was measured in ECM of donor-matched ectocervical explants incubated with culture medium (CM), seminal plasma (SP) 25% or SP50%, in the presence or absence of indomethacin 10μM, for 2, 4 and 12 h. On the right, cy...

Gene expression in ectocervical tissue explants. N-fold change in gene expression measured in ectocervical explants incubated with seminal plasma (SP) 50% and/or indomethacin 10μM for 4 and 12 h, compared to donor-matched untreated explants (CM). Values are reported as median and IQR (n = 7). P<0.05 (in bold) indicates a statistically significant d...

Document S1. Figures S1–S6, Tables S1 and S4, and Supplemental Experimental Procedures

Tissue-resident memory T (Trm) cells form a heterogeneous population that provides localized protection against pathogens. Here, we identify CD49a as a marker that differentiates CD8⁺ Trm cells on a compartmental and functional basis. In human skin epithelia, CD8⁺CD49a⁺ Trm cells produced interferon-γ, whereas CD8⁺CD49a⁻ Trm cells produced interleu...

Blood pressure regulation is known to be maintained by a neuro-endocrine circuit, but whether immune cells contribute to blood pressure homeostasis has not been determined. We previously showed that CD4(+) T lymphocytes that express choline acetyltransferase (ChAT), which catalyzes the synthesis of the vasorelaxant acetylcholine, relay neural signa...

Invariant NKT (iNKT) cells are innate-like T cells that respond rapidly with a broad range of effector functions upon recognition of glycolipid Ags presented by CD1d. HIV-1 carries Nef- and Vpu-dependent mechanisms to interfere with CD1d surface expression, indirectly suggesting a role for iNKT cells in control of HIV-1 infection. In this study, we...

The female genital tract (FGT) mucosa is a critically important site for immune defense against microbes. Mucosal-associated invariant T (MAIT) cells are an innate-like T-cell population that recognizes microbial riboflavin metabolite antigens in an MR1-dependent manner. The role of MAIT cells in the FGT mucosa is unknown. Here, we found that MAIT...

Supplementary Figure 1. (a) Representative plot showing the co-expression of CD161 on CD45+IL-18Rα+Vα7.2+ and co-expression of IL-18Rα on CD45+CD3+CD161highVα7.2+ (MAIT) cells obtained from cervix. (b) Representative histograms of one of three replicate experiments showing the expression levels (GMFI, geometric mean intensity fluorescence) of IL-18...

Human tissue explants are a valuable tool to study the interactions between host and infectious agents. They reliably mimic many important aspects of tissue cytoarchitecture and functions and allow us the investigation of the mechanisms of microbial pathogenesis under controlled laboratory conditions. One of the advantages of this system is that, u...

The effects of sex hormones on the immune defenses of the female genital mucosa and its susceptibility to infections are poorly understood. The injectable hormonal contraceptive depot medroxyprogesterone acetate (DMPA) may increase risk of HIV-1 acquisition. We assessed the local concentration in the female genital mucosa of cationic polypeptides w...

Currently, whether hormonal contraceptives affect male to female human immunodeficiency virus (HIV) transmission is debated. In this study, we investigated whether the use of progesterone-based intrauterine devices (pIUD) is associated with a thinning effect on the ectocervical squamous epithelium, down-regulation of epithelial junction proteins, a...

ProblemSex hormones can influence the immune defenses of the female genital tract (FGT) and its susceptibility to infections. Here we investigated the effect of different hormonal contraceptives on the production of antimicrobial peptides (AMPs) in different compartments of the female genital mucosa (FGM), secretions and tissue.Method of studyCervi...

We herein report the application of the phosphorodiamidate phosphate prodrug approach to a series of thirteen nucleoside analogs with antiviral or anticancer activity. Twenty-five symmetrical phosphorodiamidates were synthesized, bearing esterified l-Alanine (and in one case d-Alanine) in the prodrug moiety, each as single stereoisomer. The presenc...

Overview of the cytokine profiles of PHA-stimulated PBMC incubated for 24 h with medium only or with 500 µg/ml of PMEO-DAPym, adefovir or tenofovir. Supernatants from PHA-stimulated PBMC cultures derived from healthy donors were collected and cytokine levels were measured by the Bio-Plex array system. (TIF)

Overview of the cytokine profiles of PBMC incubated for 24 h with medium only or with 500 µg/ml of PMEO-DAPym, adefovir or tenofovir. Supernatants from PBMC cultures derived from healthy donors were collected and cytokine levels were measured by the Bio-Plex array system. (TIF)

Human immunodeficiency virus (HIV) infection is often accompanied by infection with other pathogens, in particular herpes simplex virus type 2 (HSV-2). The resulting coinfection is involved in a vicious circle of mutual facilitations. Therefore, an important task is to develop a compound that is highly potent against both viruses to suppress their...

The majority of HIV-1 infections in women transmit through vaginal intercourse, in which virus-containing semen is deposited on the cervico-vaginal mucosa. Semen is more than a mere carrier of HIV-1, since it contains many biological factors, in particular cytokines, that may affect HIV-1 transmission. The concentration of interleukin (IL)-7, one o...

The majority of HIV-1 infections in women occur through vaginal intercourse, in which virus-containing semen is deposited on the cervico-vaginal mucosa. Semen is more than a mere carrier of HIV-1, since it contains many biological factors, in particular cytokines, that may affect HIV-1 transmission. The concentration of interleukin (IL)-7, one of t...

Background. Interleukin (IL)-7 is one of the most prominent cytokines in semen of healthy fertile men and its concentration is increased in semen of HIV-1-infected individuals. IL-7 plays a central role in T cell development and homeostasis, however, despite evidence of elevated IL-7 levels in semen, little is known on its role in HIV-1 transmissio...

Although it is established that the levels of individual cytokines are altered by HIV-1 infection, the changes in cytokine interrelations that organize them into networks have been poorly studied. Here, we evaluated these networks in HIV-infected and HIV-uninfected individuals in fluid compartments that are critical for HIV-1 pathogenesis and trans...

PROBLEM: Although it is established that the levels of individual cytokines are altered by HIV-1 infection, the changes in cytokine interrelations that organize them into networks have been poorly studied. Here, we evaluated these networks in HIV-infected and HIV-uninfected individuals in fluid compartments that are critical for HIV-1 pathogenesis...

Multiple clinical trials have demonstrated that herpes simplex virus 2 (HSV-2) suppressive therapy using acyclovir (ACV) or valacyclovir in HIV-1/HSV-2-infected persons increased the patient's survival and decreased the HIV-1 load. It has been shown that the incorporation of ACV-monophosphate into the nascent DNA chain instead of dGMP results in th...

Semen is the main carrier of sexually transmitted viruses, including human immunodeficiency virus type 1 (HIV-1). However, semen is not just a mere passive transporter of virions but also plays an active role in HIV-1 transmission through cytokines and other biological factors. To study the relationship between viruses and the chemokine-cytokine ne...

The HIV reverse-transcriptase inhibitor, tenofovir, was recently formulated into a vaginal gel for use as a microbicide. In human trials, a 1% tenofovir gel inhibited HIV sexual transmission by 39% and, surprisingly, herpes simplex virus-2 (HSV-2) transmission by 51%. We demonstrate that the concentration achieved intravaginally with a 1% tenofovir...

Citation Merbah M, Introini A, Fitzgerald W, Grivel J-C, Lisco A, Vanpouille C, Margolis L. Cervico-vaginal tissue ex vivo as a model to study early events in HIV-1 infection. Am J Reprod Immunol 2011; 65: 268–278 Vaginal intercourse remains the most prevalent route of infection of women. In spite of many efforts, the detailed mechanisms of HIV-1 t...