Evolution of the Cutaneous Immune Response to Experimental Haemophilus ducreyi Infection and Its Relevance to HIV-1 Acquisition This information is current as of September 25, 2021. Tricia L. Humphreys, Carol T. Schnizlein-Bick, Barry P. Katz, Lee Ann Baldridge, Antoinette F. Hood, Robert A. Hromas and Stanley M. Spinola J Immunol 2002; 169:6316-6323; ; doi: 10.4049/jimmunol.169.11.6316 Downloaded from http://www.jimmunol.org/content/169/11/6316 References This article cites 57 articles, 19 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/169/11/6316.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 25, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Evolution of the Cutaneous Immune Response to Experimental Haemophilus ducreyi Infection and Its Relevance to HIV-1 Acquisition1,2 Tricia L. Humphreys,3* Carol T. Schnizlein-Bick,* Barry P. Katz,* Lee Ann Baldridge,† Antoinette F. Hood,‡ Robert A. Hromas,*§ and Stanley M. Spinola*¶ͦͦ Haemophilus ducreyi causes the sexually transmitted disease chancroid, which facilitates HIV-1 transmission. Skin biopsies were obtained from subjects experimentally infected with H. ducreyi to study the evolution of the immune response and immunophe- notypes relevant to transmission of HIV-1. Compared with peripheral blood, there was an enrichment of T cells and macrophages after 48 h of infection in the skin. Neutrophils became the predominant cell type by 7–9 days. By immunohistochemistry, mac- rophage-inflammatory protein-1␣ was not present early in infection, but was abundant at later stages. RANTES was present throughout the papular and pustular stages of experimental infection, but not present in uninfected control skin. Stromal cell- Downloaded from derived factor-1 was present at low levels in all samples examined. Macrophages in lesions had significantly increased expression of CCR5 and CXCR4 compared with peripheral blood cells, and CD4 T cells had significant up-regulation of CCR5. The mag- nitude of increased expression of these receptors was not replicated when PBMCs were incubated with H. ducreyi or H. ducreyi lipooligosaccharide in vitro. Together with the disruption of mucosal and skin barriers, the presence of cells with up-regulated HIV-1 coreceptors in H. ducreyi-infected lesions may provide an environment that facilitates the acquisition of R5 (CCR5), X4 http://www.jimmunol.org/ (CXCR4), and dual-tropic HIV-1 strains. The Journal of Immunology, 2002, 169: 6316–6323. aemophilus ducreyi is the etiologic agent of chancroid, a at which time patients generally seek medical attention. Because genital ulcer disease (GUD).4 Like other GUDs, chan- patients wait until the ulcerative stage to seek treatment, little is H croid increases acquisition of HIV-1 (1, 2). Chancroid is known about the initial stages of disease. endemic in certain areas of Africa and Asia, accounting for up to To characterize the immune response to H. ducreyi infection and 56% of GUD (3–8), and many chancroid endemic countries have study the pathogenesis of chancroid, we developed a human model HIV-1 seroprevalence rates exceeding 8% (9). GUDs may enhance of experimental infection that closely mimics the papular and pus- HIV-1 transmission by up to 100-fold per sexual contact, although tular stages of natural chancroid (13–15). In the model, the bacteria by guest on September 25, 2021 the exact magnitude of this cofactor effect is difficult to estimate are delivered to the epidermis and dermis on the upper arm with an (10–12). Simulation models predict that GUD had a major effect allergy testing device. One of the strengths of the model is infec- on the spread of HIV-1 in Africa in the 1990s, accounting for the tion of a relevant site of infection, human skin. The histopathology acquisition of up to 40–80% of HIV-1 infections (10). of experimental infection closely resembles that of naturally oc- In naturally occurring chancroid, H. ducreyi most likely enters curring disease (16–18). In the human model, an estimated deliv- the skin through breaks in the epithelium during intercourse. H. ered dose as low as 1–2 CFU may result in papule formation. In ducreyi infects both genital and nongenital skin, as well as mucosal contrast, animal models require inoculation with 104-107 CFU to surfaces and regional lymph nodes. Papules appear first, followed in 2–3 days by pustules. The pustules ulcerate after several weeks, establish infection (19–21). In the human model, sites may progress into pustules or resolve spontaneously. The infection is terminated after 14 days or when a painful pustule develops, gen- Departments of *Medicine, †Anatomic Pathology, ‡Dermatology, §Biochemistry, erally 7–9 days after inoculation. For subject safety, infection is ¶ ͉͉ Microbiology and Immunology, and Pathology and Laboratory Medicine, Indiana not allowed to progress to ulcers. The model is useful in that it University, Indianapolis, IN 46202 mimics the first 2 wk of an infection that in nature is probably Received for publication July 19, 2002. Accepted for publication October 1, 2002. present for 3–6 wk before patients with ulcers seek treatment (22). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance The local immune response to experimental infection consists of with 18 U.S.C. Section 1734 solely to indicate this fact. epidermal pustules filled with polymorphonuclear leukocytes and a 1 This work was supported by Grants AI31494 and AI27863 from the National In- perivascular infiltrate of T cells, macrophages, and a few B cells in stitutes of Allergy and Infectious Diseases. T.L.H. was supported by T32AI07367 from the National Institutes of Allergy and Infectious Diseases. The human challenge the dermis (14, 18). By immunohistopathologic analysis, the ma- trials were also supported by Grant MO1RR00750 to the General Clinical Research jority of T cells are CD4, CD45RO cells. In experimental infec- Center at Indiana University. tion, H. ducreyi associates with the polymorphonuclear leukocytes 2 Portions of this work were presented in abstract form at the International Congress in the pustule and with macrophages at the base of the pustule, but of Sexually Transmitted Disease in Berlin, Germany, 2001, and at the American Society for Microbiology meeting in Salt Lake City, UT, 2002. is not taken up by the phagocytes (23). By escaping phagocytic 3 Address correspondence and reprint requests to Dr. Tricia L. Humphreys, Indiana killing, the bacteria sustain the immune response that most likely University School of Medicine, 435 Emerson Hall, 545 Barnhill Drive, Indianapolis, damages the skin (22). Although the immunohistopathology of IN 46202. E-mail address: [email protected] experimental and natural infection is well described, no quantita- 4 Abbreviations used in this paper: GUD, genital ulcer disease; FSC, forward scatter; LOS, lipooligosaccharide; MIP-1␣, macrophage-inflammatory protein 1␣; SDF-1, tive studies that address kinetics of cell recruitment have been stromal cell-derived factor 1; SSC, side scatter. performed. Copyright © 2002 by The American Association of Immunologists, Inc. 0022-1767/02/$02.00 The Journal of Immunology 6317 HIV-1 is primarily sexually transmitted. HIV-1 uses CD4 as a populations of macrophages and T cells and later evolves into receptor for entry into cells, along with the coreceptors CCR5 and polymorphonuclear leukocyte predominance. We also show that CXCR4 (24, 25). HIV-1 has been classified according to corecep- HIV-1 coreceptors CCR5 and CXCR4 are enriched on macro- tor use. R5 viruses use CCR5 as a coreceptor for entry, while X4 phages, and CCR5 is enriched on T cells throughout experimental viruses use CXCR4 as a coreceptor (26). Some primary isolates of infection. HIV-1 can use both coreceptors, while many laboratory-adapted strains use CXCR4 only. Although the type of virus that is trans- Materials and Methods mitted has not been directly studied, the majority of virus in semen Human subjects are R5 (27). Shortly after seroconversion, most individuals have Twenty-two healthy adult volunteers donated 37 tissue specimens for this R5 virus circulating in their peripheral blood, although some in- study. Biopsies designated CS were obtained from seven subjects who dividuals have X4 virus or both types (28). Individuals with a participated in a previous human challenge trial (15, 18). Tissue specimens homozygous ⌬32 mutation in CCR5 are resistant to infection with for FACS analysis were obtained from 10 volunteers (189 and 195–204) HIV-1, confirming the importance of R5 viruses in transmission who were infected for the purposes of this study, 2 subjects who partici- (29–32). To our knowledge, no data exist on the type of virus pated in a mutant parent trial (186 and 190) (33), and 2 subjects (164RR and 172RR) who participated in a reinfection trial (Table I). One unin- acquired by persons who are infected with other sexually trans- fected volunteer donated normal skin (18). Informed consent was obtained mitted diseases. from the subjects in accordance with the guidelines for human experimen- GUDs such as chancroid are thought to facilitate HIV-1 acqui- tation of the U.S.