Christen G. Press

Spontaneous Resolution of Genital Chlamydia trachomatis Infection in Women and Protection from Reinfection

The natural history of chlamydia is variable and may include persisting asymptomatic infection, complications, or spontaneous resolution before treatment. Reinfection is common. We evaluated whether spontaneous resolution was associated with decreased reinfection in women returning for treatment of a positive chlamydia screening test. At enrollment, participants were tested for chlamydia, treated with azithromycin, and scheduled for a 6-month follow-up visit for repeat testing. Two hundred participants returned 1 to 12 months after treatment. Spontaneous resolution at enrollment was demonstrated in 44 (22.0%). Reinfection at follow-up occurred in 33 (16.5%), being more frequent in those with persisting infection at enrollment versus spontaneous resolution (31 of 156 [19.9%] vs 2 of 44 [4.5%]; P = .016). Adjusting for age, the odds of reinfection was 4 times higher for participants with persisting infection at enrollment (odds ratio 4.0, 95% confidence interval, 1.1-25.6; P = .034). Chlamydia treatment may attenuate protective immunity in some patients.

Investigating the Epidemiology of Repeat Chlamydia trachomatis Detection after Treatment by Using C. trachomatis OmpA Genotyping

ABSTRACT Repeat Chlamydia trachomatis detection frequently occurs within months after C. trachomatis infection treatment. The origins of such infection (persistence versus reinfection from untreated or new partners) are varied and difficult to determine. C. trachomatis strains can be differentiated by sequencing the ompA gene encoding the outer membrane protein A (OmpA). We used OmpA genotyping to investigate the epidemiology of repeat C. trachomatis detection after treatment in C. trachomatis-infected subjects seen at a sexually transmitted diseases clinic. Subjects were enrolled, tested for C. trachomatis, treated with azithromycin, and scheduled for a 6-month follow-up for repeat C. trachomatis testing. OmpA genotyping was performed on C. trachomatis-positive urogenital specimens obtained from patients at enrollment and follow-up. The enrollment visit OmpA genotypes for C. trachomatis were determined for 162 subjects (92% female, 94% African American). C. trachomatis was detected at follow-up in 39 subjects (24%). The OmpA genotype distribution at enrollment did not differ in those with versus those without repeat C. trachomatis detection. Of the 35 subjects with C. trachomatis strains genotyped at enrollment and follow-up, 7 (20%) had the same ompA sequence at both visits, while 28 (80%) had discordant sequences. A new sexual partner was reported more often in subjects with discordant C. trachomatis strains than in those with concordant strains (13 [46%] versus 1 [14%]; P = 0.195). Half of the subjects with discordant C. trachomatis strains who reported sexual activity since treatment denied a new sexual partner; 62% of these subjects reported that their partner was treated. Our study demonstrates that most repeat C. trachomatis detections after treatment were new infections with a different C. trachomatis strain rather than reinfection with the same strain. OmpA genotyping can be a useful tool in understanding the origins of repeat C. trachomatis detection after treatment.

The Predominant CD4+ Th1 Cytokine Elicited to Chlamydia trachomatis Infection in Women is Tumor Necrosis Factor-Alpha, not Interferon-Gamma.

ABSTRACT Chlamydia trachomatis infection is the most prevalent bacterial sexually transmitted infection and can cause significant reproductive morbidity in women. There is insufficient knowledge of C. trachomatis-specific immune responses in humans, which could be important in guiding vaccine development efforts. In contrast, murine models have clearly demonstrated the essential role of T helper type 1 (Th1) cells, especially interferon gamma (IFN-γ)-producing CD4+ T cells, in protective immunity to chlamydia. To determine the frequency and magnitude of Th1 cytokine responses elicited to C. trachomatis infection in humans, we stimulated peripheral blood mononuclear cells from 90 chlamydia-infected women with C. trachomatis elementary bodies, Pgp3, and major outer membrane protein and measured IFN-γ-, tumor necrosis factor alpha (TNF-α)-, and interleukin-2 (IL-2)-producing CD4+ and CD8+ T-cell responses using intracellular cytokine staining. The majority of chlamydia-infected women elicited CD4+ TNF-α responses, with frequency and magnitude varying significantly depending on the C. trachomatis antigen used. CD4+ IFN-γ and IL-2 responses occurred infrequently, as did production of any of the three cytokines by CD8+ T cells. About one-third of TNF-α-producing CD4+ T cells coproduced IFN-γ or IL-2. In summary, the predominant Th1 cytokine response elicited to C. trachomatis infection in women was a CD4+ TNF-α response, not CD4+ IFN-γ, and a subset of the CD4+ TNF-α-positive cells produced a second Th1 cytokine.

O01.1 Association of Genetic Variants with Chlamydia Trachomatis Reinfection

Background Up to 20% of Chlamydia trachomatis (CT)-infected patients are reinfected within months after treatment, suggesting some fail to develop protective immunity. Genetic determinants influencing CT reinfection risk have not been fully elucidated. Our primary research objective is to identify genetic determinants of CT reinfection. Based on previously reported associations of HLA class II alleles with CT complications, our initial investigations focus on HLA class II genes. Methods In an ongoing prospective natural history study, CT-infected subjects are enrolled, treated with azithromycin 1 g single dose, and return for a 6-month follow-up visit for repeat CT testing using the Gen-Probe APTIMA Combo 2 assay (Gen-Probe, Inc., San Diego, CA). HLA class II alleles are resolved by a combination of PCR-based techniques. Genomic DNA is stored for further genotyping. Results A total of 199 African American subjects have been studied to date: 90% women and median age 23. CT reinfection at follow-up was noted in 18%. Subjects with HLA-DQB1*05 more often had reinfection (20 [26%] vs. 16 [13%], P = 0.018), which remained significant after controlling for age and gender (OR 2.6, 95% CI 1.2–5.6, P = 0.012). Other HLA-DQB1 alleles were not significantly associated with reinfection (P ≥ 0.1). Conclusion HLA-DQB1* 05was associated with CT reinfection, suggesting it could influence protective immunity. More comprehensive genotyping from larger prospectively studied cohorts should help confirm or refine this observation. Analysis of additional HLA class II genes and genes beyond the human MHC is in progress.

HLA-DQB1*06 is a risk marker for chlamydia reinfection in African American women

Associations between human leukocyte antigen (HLA) variants and chlamydia-related outcomes have been inconsistent. We previously identified HLA-DQB1*06 as a risk marker for chlamydia reinfection in a cohort of predominately HIV-infected adolescents. As chlamydia reinfection can lead to reproductive complications, validation of this finding in HIV-seronegative women may help reveal the underlying biology. We performed HLA-DQB1 genotyping in HIV-seronegative, chlamydia-infected African American women who were evaluated for reinfection at 3- and 6-month visits after treatment. Of 185 evaluable women for whom HLA-DQB1 genotyping was performed, only HLA-DQB1*06 was associated with chlamydia reinfection (Pu2009=u20090.009), with no evidence of a dose–response effect for this allele. African American women with HLA-DQB1*06 may warrant more frequent chlamydia screening. More comprehensive genotyping of HLA class II and neighboring genes is needed to establish whether HLA-DQB1*06 is a causal variant for chlamydia reinfection or a surrogate for other causal variants in the major histocompatibility complex.

An Adaptive Chlamydia trachomatis-Specific IFN-γ-Producing CD4+ T Cell Response Is Associated With Protection Against Chlamydia Reinfection in Women

Background: Adaptive immune responses that mediate protection against Chlamydia trachomatis (CT) remain poorly defined in humans. Animal chlamydia models have demonstrated that CD4+ Th1 cytokine responses mediate protective immunity against reinfection. To better understand protective immunity to CT in humans, we investigated whether select CT-specific CD4+ Th1 and CD8+ T cell cytokine responses were associated with protection against CT reinfection in women. Methods: Peripheral blood mononuclear cells were collected from 135 CT-infected women at treatment and follow-up visits and stimulated with CT antigens. CD4+ and CD8+ T-cells expressing IFN-γ, TNF-α, and/or IL-2 were assessed using intracellular cytokine staining and cytokine responses were compared between visits and between women with vs. without CT reinfection at follow-up. Results: A CD4+TNF-α response was detected in the majority (77%) of study participants at the treatment visit, but a lower proportion had this response at follow-up (62%). CD4+ IFN-γ and CD4+ IL-2 responses occurred less frequently at the treatment visit (32 and 18%, respectively), but increased at follow-up (51 and 41%, respectively). CD8+ IFN-γ and CD8+ TNF-α responses were detected more often at follow-up (59% for both responses) compared to the treatment visit (30% for both responses). At follow-up, a CD4+IFN-γ response was detected more often in women without vs. with reinfection (60 vs. 33%, P = 0.005). Conclusions: Our findings suggest that a CT-specific CD4+ IFN-γ response is associated with protective immunity against CT reinfection and is thus an important component of adaptive immunity to CT in women.

Lower Levels of Cervicovaginal Tryptophan are Associated with Natural Clearance of Chlamydia in Women.

Chlamydiatrachomatis (Ct) infection causes significant morbidity. In vitro studies demonstrate that Ct growth inhibition occurs by interferon-gamma (IFN-γ)-mediated depletion of intracellular tryptophan, and some Ct strains utilize extracellular indole to restore tryptophan levels. Whether tryptophan levels are associated with Ct infection clearance in humans remains unknown. We evaluated tryptophan, indole, and IFN-γ levels in cervicovaginal lavages from women with either naturally cleared or persisting Ct infection. Women who cleared infection had significantly lower tryptophan levels and trended toward lower IFN-γ levels compared to women with persisting infection. Due to its volatility, indole was not measurable in either group.

T cell phenotypes in women with Chlamydia trachomatis infection and influence of treatment on phenotype distributions

T cell phenotypes involved in the immune response to Chlamydia trachomatis (CT) have not been fully elucidated in humans. We evaluated differences in T cell phenotypes between CT-infected women and CT-seronegative controls and investigated changes in T cell phenotype distributions after CT treatment and their association with reinfection. We found a higher expression of T cell activation markers (CD38+HLA-DR+), T helper type 1 (Th1)- and Th2-associated effector phenotypes (CXCR3+CCR5+ and CCR4+, respectively), and T cell homing marker (CCR7) for both CD4+ and CD8+ T cells in CT-infected women. At follow-up after treatment of infected women, there were a lower proportion of CD4+ and CD8+ T cells expressing these markers. These findings suggest a dynamic interplay of CD4+ and CD8+ T cells in CT infection, and once the infection is treated, these cell markers return to basal expression levels. In women without reinfection, a significantly higher proportion of CD8+ T cells co-expressing CXCR3 with CCR5 or CCR4 at follow-up was detected compared to women with reinfection, suggesting they might play some role in adaptive immunity. Our study elucidated changes in T cell phenotypes during CT infection and after treatment, broadening our understanding of adaptive immune mechanisms in human CT infections.

P3.258 Investigating the Epidemiology of Repeat Chlamydia Trachomatis Detection After Treatment Using Chlamydia Trachomatis Omp A Genotyping

Background Detection of Chlamydia trachomatis (CT) infection within months of initial diagnosis and treatment is a common occurrence. Origins of such infection (persistence vs. reinfection from an untreated or a new partner) are complex. CT strains can be differentiated by complete nucleotide sequence analysis of the ompA gene, encoding an antigenically diverse surface protein outer membrane protein A (OmpA). We are evaluating urogenital CT OmpA genotypes in an ongoing prospective CT natural history study in order to investigate the epidemiology of repeat CT detection after treatment. Methods CT-infected subjects are prospectively enrolled, treated with azithromycin, and return for a 6-month follow-up visit for repeat CT testing using the Gen-Probe APTIMA Combo 2 (Gen-Probe, Inc., San Diego, CA). Urogenital specimens are collected at enrollment and follow-up, from which CT strains are genotyped by ompA amplification and sequencing. Results Enrollment visit genotypes have been determined for 145 subjects to date (91% female, 93% African American). CT infection was detected at follow-up in 39 (27%). Enrollment genotype distribution did not significantly differ in those without versus with repeat CT detection at follow-up (major genotypes: D/Da 25%,23%; E 22%,28%; F 13%,15%; I/Ia 17%,15%; J/Ja 12%,13%). Of 35 subjects with CT strains genotyped from both enrollment and follow-up visits, 7 (20%) had the same CT strain at both visits versus 28 (80%) with a different strain at follow-up. Sexual activity post-treatment was reported in 32 subjects with strains genotyped at both visits; a new sexual partner was reported more often in subjects with discordant vs. concordant strains (52% vs. 14%, p = 0.1). Conclusion Baseline CT Omp A genotype did not predict repeat CT detection. Most repeat CT infection detections were new infections with a different CT strain. Genotyping will be a useful tool in understanding the origins of repeat CT infection detection after treatment.