Herpes Simplex Virus Type 2 Co-infection Research Articles

Cervical Cytology and Herpes Simplex Virus Type-2 Serology Among Human Immune Deficiency Virus Infected Women on Highly Active Antiretroviral Therapy in Enugu, Nigeria.

The prevalence of Human Immunodeficiency Virus (HIV) and Herpes Simplex Virus type-2 (HSV-2) infections are high and the programme for Cervical Cancer Screening is weak in Nigeria. Prevalence of Herpes Simplex Virus Type-2 co-infection and cervical cytology among HSV-2 co-infected Human Immunodeficiency Virus Sero-positive (HIV+) women on Highly Active Antiretroviral Therapy (HAART) attending Human Immunodeficiency Virus clinic at University of Nigeria Teaching Hospital (UNTH) Ituku/Ozalla Enugu, Nigeria. A cross-sectional, hospital-based study. Active participants included 105 HIV seropositive women on HAART and104 HIV seronegative (HIV-) women who passed inclusion criteria and signed written informed consent. Each participant was coded with a specific number. A structured questionnaire was used to obtain the socio-demographic and medical history. Serum was obtained for HSV-2 serology test for all participants and HIV screening for HIV-negative participants. Cervical smears were collected for Papanicolaou stains and Immunocytochemistry using anti-P16INKa antibody. Prevalence of HSV-2+ was 50.5% among HIV+ women on HAART and 16.3% among HIV- women, Odds Ratios [95% CI]; p-value was 5.21 [2.74-9.94]; p < 0.0001. HIV+ women on HAART co-infected with HSV-2 significantly had more Cervical Lesions, 11.4% compared to HIVwomen uninfected with HSV-2, 4.8%, OR [95% CI]; p-value 4.8 (1.58-14.54); p = 0.006. The prevalence of HSV-2 was significantly high among HIV+ women on HAART. HSV-2+ co-infection could be an enhancer of Cervical Lesions among HIV+ women on HAART. Hence, anti-herpetic agent introduction and screening for HSV-2 among HIV+ patients are recommended.

Altered HIV-1 Viral Copy Number and Gene Expression Profiles of Peripheral (CEM CCR5+) and Mucosal (A3R5.7) T Cell Lines Co-Infected with HSV-2 In Vitro.

Co-infecting pathogens have been speculated to influence Human Immunodeficiency Virus (HIV) disease progression. Herpes Simplex Virus Type-2 (HSV-2), another sexually transmitted pathogen, is commonly observed in individuals with HIV-1. Some clinical studies have observed an increase in HIV-1 viral copy number in HSV-2 co-infected individuals. In vitro studies have also demonstrated an increase in the expression of HIV-1 co-receptors on immune cells infected with HSV-2. Although both the viruses show distinctive persistent infection, the influence of HSV-2 on HIV-1 is poorly understood. Here we present a comparative analysis of primary CD4+ T-cells and four different T-cell lines (PM-1, CEM CCR5+, MOLT4 CCR5+, and A3R5.7) to assess the influence of HSV-2 co-infection on HIV-1 replication in vitro. Cell lines indicating significant changes in HIV-1 viral copy number [CEM CCR5+ (0.61 Log10), A3R5.7 (0.78 Log10)] were further evaluated for the infectivity of HIV-1 virions and the changes in gene expression profiles of HSV-2/HIV-1 co-infected and mono-infected cells, which were further confirmed by qPCR. Significant changes in NUP, MED, and VPS mRNA expression were observed in the gene expression profiles in co-infected CEM CCR5+ and A3R5.7 cells. In both cell lines, it was observed that the WNT signaling, PI3 kinase, apoptosis, and T-cell activation pathways were negatively affected in co-infected cells. The data suggest that HSV-2 infection of T-cells may influence the expression of genes that have been previously shown to affect HIV-1 replication in vitro. This idea needs to be explored further to identify anti-viral targets for HSV-2 and HIV-1.

Seroprevalence of Herpes Simplex Virus Type 2 and HIV Co-Infection Among Pregnant Women in Jigawa State

Herpes simplex virus type 2 (HSV-2) infections is one of the most common sexually transmitted infections Worldwide. It facilitates the acquisition of HIV and is the primary cause of genital herpes which when acquired by women during pregnancy account for half of the mobidity and mortality among neonates. Lifelong latent HSV-2 infection raises concern among women of reproductive age considering the risk of neonatal transmission. In Nigeria screening for HSV-2 and co-infection with HIV in antenatal clinics is not routinely done. A cross-sectional study, was carried out among pregnant women attending antenatal clinics in four hospitals in Jigawa state. A total of 300 consenting pregnant women were enrolled, the study involved collection of sociodemographic data and laboratory determination of HSV-2 immunoglobulin G (IgG) and HIV seroprevalence using WHO standard procedure of Enzyme Linked Immunosorbent Assay (ELISA) test kit (DIAPRO Diagnostic Bioprobes, Milano, Italy), UNI-GOLD Wicklow, Ireland and ALERE Determine, respectively. Statistical significance was determined at p<0.05Of the 300 samples tested, overall seroprevalence of 55 (18.3%) and 44 (14.7%) for HSV-2 and HIV were recorded. The HSV-2 and HIV Co-infection rate was 16 (5.3%) among respondent. About 87.5% of the co-infection occurred in respondents that were in their 2^nd Trimester. Addionally, co-infection was found to be high among respondents between the age of 15-34yrs with 81.3%. In this study, education status, parity, stage of pregnancy, occupation, History of blood transfusion had no statistical significance with HSV-2 and HIV co-infection (p>0.05). This study has demonstrated the existence and risk of neonatal herpes in the study area. It is recommended that HSV testing should be placed among the standard medical checkup tests in the area, again advocacy and public awareness on the potential public health burden of HSV-2 and HIV co-infection should be observed.

Progesterone augments cell susceptibility to HIV-1 and HIV-1/HSV-2 co-infections.

In human immunodeficiency virus type 1 (HIV-1)-infected women, oral or injectable progesterone containing contraceptive pills may enhance HIV-1 acquisition in vivo, and the mechanism by which this occurs is not fully understood. In developing countries, Herpes simplex virus type-2 (HSV-2) co-infection has been shown to be a risk for increase of HIV-1 acquisition and, if co-infected women use progesterone pills, infections may increase several fold. In this study, we used an in vitro cell culture system to study the effects of progesterone on HIV-1 replication and to explore the molecular mechanism of progesterone effects on infected cells. In our in vitro model, CEMss cells (lymphoblastoid cell line) were infected with either HIV-1 alone or co-infected with HSV-2. HIV-1 viral load was measured with and without sex hormone treatment. Progesterone-treated cells showed an increase in HIV-1 viral load (1411.2 pg/mL) compared with cells without progesterone treatment (993.1 pg/mL). Increased cell death was noted with HSV-2 co-infection and in progesterone-treated cells. Similar observations were noted in peripheral blood mononuclear cells (PBMC) cells derived from three female donors. Progesterone-treated cells also showed reduced antiviral efficacy. Inflammatory cytokines and associations with biomarkers of disease progression were explored. Progesterone upregulated inflammatory cytokines and chemokines conversely and downregulated anti-apoptotic Bcl-2 expression. Nuclear protein analysis by electrophoretic mobility shift assay showed the association of progesterone with progesterone response element (PRE), which may lead to downregulation of Bcl-2. These data indicate that progesterone treatment enhances HIV-1 replication in infected cells and co-infection with HSV-2 may further fuel this process.

Parallel declines in HIV and hepatitis C virus prevalence, but not in herpes simplex virus type 2 infection: A 10-year, serial cross-sectional study in an inner-city emergency department

BackgroundThe Johns Hopkins Hospital Emergency Department (JHHED) has served as an observational window on the HIV epidemic in a socioeconomically depressed, urban population. We previously reported that HIV incidence among JHHED patients is decreasing and that prevalence has declined from 11.4% in 2003–5.6% in 2013. ObjectivesThis study sought to observe temporal trends in hepatitis C virus (HCV) and herpes simplex virus type 2 (HSV-2) seroprevalence, which are surrogate markers for parenteral and sexual risk behavior, respectively. Study designIdentity unlinked-serosurveys were conducted over 6–8 weeks in the adult JHHED in 2003, 2007, and 2013. Excess sera from 10,274 patients, previously tested for HIV, were assayed for HSV-2 and HCV antibodies. ResultsOverall HCV seroprevalence declined steadily from 22.0% in 2003–13.8% in 2013 (Ptrend<0.01), and was significant by all gender and race strata. Overall HSV-2 prevalence declined from 55.3% in 2003–50.0% in 2013 (Ptrend<0.01), but was non-significant after adjustment for demographics. Among HIV+ individuals<45years of age, there was a significant decrease in the proportion of individuals with HCV co-infection [without HSV-2] (Ptrend=0.02) from 2003 to 2013, however, there was an increase in individuals with HSV-2 co-infection [without HCV] (Ptrend < 0.01). DiscussionLittle change in age-specific HSV-2 prevalence suggests the decrease in HIV prevalence was likely not associated with changes in sexual risk behavior. In addition to clinical interventions, strategies to address sexual health disparities and continued parenteral harm-reduction efforts are needed to further drive the decline in HIV.

Herpes Simplex Virus: The Interplay Between HSV, Host, and HIV-1.

Herpes simplex virus proteins interact with host (human) proteins and create an environment conducive for its replication. Genital ulceration due to herpes simplex virus type 2 (HSV-2) infections is an important clinical manifestation reported to increase the risk of human immunodeficiency virus type 1 (HIV-1) acquisition and replication in HIV-1/HSV-2 coinfection. Dampening the innate and adaptive immune responses of the skin-resident dendritic cells, HSV-2 not only helps itself, but creates a "yellow brick road" for one of the most dreaded viruses HIV, which is transmitted mainly through the sexual route. Although, data from clinical trials show that HSV-2 suppression reduces HIV-1 viral load, there are hardly any reports presenting conclusive evidence on the impact of HSV-2 coinfection on HIV-1 disease progression. Be that as it may, understanding the interplay between these three characters (HSV, host, and HIV-1) is imperative. This review endeavors to collate studies on the influence of HSV-derived proteins on the host response and HIV-1 replication. Studying such complex interactions may help in designing and developing common strategies for the two viruses to keep these "partners in crime" at bay.

Joint modeling of correlated binary outcomes: HIV-1 and HSV-2 co-infection

Herpes Simplex Virus Type 2 (HSV-2) facilitates the sexual acquisition and transmission of HIV-1 infection and is highly prevalent in most regions experiencing severe HIV epidemics. In sub-Saharan Africa, where HIV infection is a public health burden, the prevalence of HSV-2 is substantially high. The high prevalence of HSV-2 and the association between HSV-2 infection and HIV-1 acquisition could play a significant role in the spread of HIV-1 in the region. The objective of our study was to identify risk factors for HSV-2 and HIV-1 infections among men in sub-Saharan Africa. We used a joint response model that accommodates the interdependence between the two infections in assessing their risk factors. Simulation studies show superiority of the joint response model compared to the traditional models which ignore the dependence between the two infections. We found higher odds of having HSV-2/HIV-1 among older men, in men who had multiple sexual partners, abused alcohol, or reported symptoms of sexually transmitted infections. These findings suggest that interventions that identify and control the risk factors of the two infections should be part of HIV-1 prevention programs in sub-Saharan Africa where antiretroviral therapy is not readily available.

Herpes simplex virus type-2 stimulates HIV-1 replication in cervical tissues: implications for HIV-1 transmission and efficacy of anti-HIV-1 microbicides.

Herpes Simplex virus Type-2 (HSV-2) increases the risk of HIV-1 acquisition, yet the mechanism for this viral pathogen to regulate the susceptibility of the cervicovaginal mucosa to HIV-1 is virtually unknown. Using ex vivo human ectocervical tissue models, we report greater levels of HIV-1 reverse transcription, DNA integration, RNA expression, and virions release in HIV-1/HSV-2 co-infected tissues compared with HIV-1 only infected tissues (P<0.05). Enhanced HIV-1 replication was associated with increased CD4, CCR5, and CD38 transcription (P<0.05) and increased number of CD4+/CCR5+/CD38+ T cells in HIV-1/HSV-2 co-infected tissues compared with tissues infected with HIV-1 alone. Tenofovir (TFV) 1% gel, the leading microbicide candidate, demonstrated only partial protection against HIV-1, when applied vaginally before and after sexual intercourse. It is possible that mucosal inflammation, in particular that induced by HSV-2 infection, may have decreased TFV efficacy. HSV-2 upregulated the number of HIV-1-infected cells and elevated the concentration of TFV needed to decrease HIV-1 infection. Similarly, only high concentrations of TFV inhibited HSV-2 replication in HIV-1/HSV-2-infected tissues. Thus, HSV-2 co-infection and mucosal immune cell activation should be taken into consideration when designing preventative strategies for sexual transmission of HIV-1.

Herpes Simplex Virus Type 2 Coinfection Does Not Accelerate CD4 Count Decline in Untreated HIV Infection

Herpes simplex virus type 2 (HSV-2) reactivations are associated with increased HIV load, but whether HSV-2 coinfection accelerates HIV disease is unclear. We compared rates of CD4 count decline according to HSV-2 status in untreated HIV-infected adults. HIV-infected patients with a past period of antiretroviral therapy (ART)-untreated follow-up with initial CD4 count of 400-900 cells/mm(3) and no chronic anti-HSV therapy were included. HSV-2 status was determined by HerpeSelect enzyme-linked immunosorbent assay. Rates of CD4 count change were compared by HSV-2 status using mixed linear regression models, and time to the first of ART initiation or CD4 <350 cells/mm(3) using proportional hazards models. Of 218 patients included, 123 (56.4%) were seropositive for HSV-2 and 161 (73.8%) for HSV-1. In univariate analysis, the difference in the rate of CD4 count change associated with HSV-2 was not statistically significant at +13.6 cells/mm(3)/year (P = .12). Results were similar at -4.5 cells/mm(3)/year (P = .68) after adjustment for sex, HSV type 1, oral and genital herpes symptoms, immigrant status, and the interaction of immigrant status with time. However, HSV-2 seropositivity was associated with a shorter time to the first of ART initiation or CD4 <350 cells/mm(3), with an adjusted hazard ratio of 2.07 (95% confidence interval, 1.28-3.33). HSV-2 coinfection was not associated with the rate of CD4 count decline during ART-untreated HIV infection, but was associated with an earlier combined endpoint of ART initiation or CD4 <350 cells/mm(3). Attenuating effects of acyclovir on HIV disease progression observed in recent clinical trials may result from direct anti-HIV activity rather than indirect benefits from HSV-2 suppression.

Serum apoptosis markers in HIV-infected patients with human herpesvirus type 8 and herpes simplex virus type 2 co-infection

This study aimed to examine the influence of human herpesvirus type 8 (HHV-8) and herpes simplex virus type 2 (HSV-2) co-infections on apoptosis serum markers in human immunodeficiency virus (HIV)-infected patients. Sera from 110 HIV-infected and 59 HIV-uninfected individuals were analyzed for soluble Fas (sFas), sFas ligand (sFasL), caspase-8, and Bcl-2. The findings of HIV-infected patients with no co-infection (n = 37), HIV-infected patients with HHV-8 co-infection (n = 22), HIV-infected patients with HSV-2 co-infection (n = 51), and patients with HSV-2 co-infection and no HIV infection (n = 20) were compared to controls (reference group) with no HIV, HSV-2, and HHV-8 co-infections (n = 39). Soluble Fas and sFasL concentrations were the highest in HIV and HHV-8 co-infected patients (medians, 912.7 pg/ml and 74.3 pg/mL, respectively). No difference in caspase-8 concentrations was found, whereas Bcl-2 concentrations were the highest in HIV and HHV-8 co-infected individuals. Older age was associated with higher sFas (p < 0.001) and lower sFasL (p = 0.04) concentrations. In a robust regression model adjusted for age, the log-transformed sFas concentrations were significantly lower in HIV-infected patients with no co-infections (β = -0.244; p < 0.001) and higher in HIV and HHV-8 co-infected patients (β = 0.216; p = 0.012) compared to the reference group. Soluble FasL was significantly lower in HIV-infected patients with no co-infections (β = -0.284; p = 0.005) and in HIV-infected patients with HSV-2 co-infection (β = -0.381; p < 0.001) compared to the reference group. Soluble FasL was also higher in HIV and HHV-8 co-infected patients compared to controls (β = 0.248; p = 0.036). Our results suggest that HHV-8 and HSV-2 may have a significant effect on Fas-FasL-mediated apoptosis in HIV-1 patients. HHV-8 upregulates while HSV-2 downregulates sFas and sFasL.

Effects of HIV-1 and Herpes Simplex Virus Type 2 Infection on Lymphocyte and Dendritic Cell Density in Adult Foreskins from Rakai, Uganda

Male circumcision reduces human immunodeficiency virus (HIV) and herpes simplex virus type 2 (HSV-2) acquisition, and HSV-2 infection is associated with an increased risk of HIV acquisition. To assess the cellular basis for these associations, we estimated immunologic cellular densities in foreskin tissue. Immunostained CD1a(+) dendritic cell and CD4(+) and CD8(+) T cell densities were quantified in foreskin samples obtained from medical circumcision in Rakai, Uganda (35 HIV-infected, HSV-2-infected men; 5 HIV-infected, HSV-2-uninfected men; 22 HIV-uninfected, HSV-2-infected men; and 29 HIV-uninfected, HSV-2-uninfected men. CD1A(+) dendritic cell densities did not vary by HIV or HSV-2 status. Compared with densities in HIV-uninfected, HSV-2-uninfected men (mean, 26.8 cells/mm(2)), CD4(+) T cell densities were similar in the HIV-infected, HSV-2-infected group (mean, 28.7 cells/mm(2)), were significantly decreased in the HIV-infected, HSV-2-uninfected group (mean, 11.2; P < .05), and were increased in the HIV-uninfected, HSV-2-infected group (mean, 68.7; P < .05). Dermal CD8(+) T cell densities were higher in the HIV and HSV-2-coinfected group (mean, 102.9) than in the HIV-uninfected, HSV-2-uninfected group (mean, 10.0; P < .001), the HIV-infected, HSV-2-uninfected group (mean, 27.3; P < .001), and the HIV-uninfected, HSV-2-infected group (mean, 25.3; P < .005). The increased CD4(+) cellular density in the HIV-uninfected, HSV-2-infected men may help to explain why HSV-2-infected men are at increased risk of HIV acquisition. The absence of this increase in men coinfected with both HIV and HSV-2 is likely in part the result of the progressive loss of CD4(+) cells in HIV infection. Conversely, HIV and HSV-2 coinfection appears to synergistically increase CD8(+) T cell densities.

Coinfection with Herpes Simplex Virus Type 2 Is Associated with Reduced HIV‐Specific T Cell Responses and Systemic Immune Activation

Chronic coinfection with herpes simplex virus type 2 (HSV-2) and human immunodeficiency virus (HIV) has been associated with an increased HIV viral load and more rapid disease progression, perhaps related to HSV-2-associated alterations in host immunity. Studies were nested within (1) a cross-sectional study of men coinfected with HIV and HSV-2 and (2) women not infected with HIV, both before and after HSV-2 acquisition. HSV-2 infection status was determined by ELISA. HIV-specific CD8(+) T cell epitopes were mapped, and proliferation of HIV-specific cells was also assessed. Systemic inflammatory and regulatory T cell populations were assayed by flow cytometry. The breadth of both the HIV-specific CD8(+) T cell interferon-gamma and proliferative responses was reduced in participants coinfected with HIV and HSV-2, independent of the HIV plasma viral load and CD4(+) T cell count, and the magnitude of the responses was also reduced. HSV-2 infection in this group was associated with increased T cell CD38 expression but not with differences in the proportion of CD4(+) FoxP3(+) regulatory T cells. However, in women not infected with HIV, acquisition of HSV-2 was associated with an increase in the proportion of regulatory T cells. HSV-2 coinfection was associated with reduced HIV-specific T cell responses and systemic inflammation. The immune effects of HSV-2 may underlie the negative impact that this coinfection has on the clinical course of HIV infection.

Maternal Herpes simplex virus type 2 infection, syphilis and risk of intra-partum transmission of HIV-1: results of a case control study

Genital ulcer disease including that caused by Herpes simplex virus type 2 (HSV-2) and syphilis facilitates sexual transmission of HIV-1. The effect of these infections on intra-partum mother-to-child-transmission (MTCT) of HIV-1 is unknown. A case-control study was conducted using archived sera from HIV-1 positive women enrolled in ZVITAMBO, an MTCT trial. Cases were 509 women who transmitted HIV-1 to their infants intra-partum; controls were 1018 women whose infants remained uninfected at 12 months. Maternal serum collected at delivery, were tested for HSV-2 antibody. The 6-week post-partum sample was also tested for syphilis by RPR and TPHA to identify women with incubating or active syphilis at delivery. Rates of prevalent and incident HSV-2 and recently acquired syphilis were compared between cases and controls. Overall prevalence of maternal HSV-2 and active syphilis at delivery were 82.5% [95% confidence interval (CI), 80.6-84.5] and 4.0% (95% CI, 3.0-5.1), respectively. Prevalent HSV-2 was associated with increased intra-partum MTCT [adjusted odds ratio (OR), 1.50; 95% CI, 1.09-2.08]. The proportion of intra-partum transmissions potentially attributable to prevalent HSV-2 infection was 28.4% (95% CI, 7.3-44.7). Maternal infection with active syphilis at delivery was not associated with intra-partum MTCT (unadjusted OR, 0.89; 95%CI, 0.49-1.59; adjusted OR, 0.64; 95% CI, 0.34-1.20). HSV-2 infection is common among HIV-1-positive women and is associated with an increased risk of intra-partum MTCT. More than 25% of intra-partum MTCT may be attributable to maternal HSV-2 co-infection. Active maternal syphilis at the time of delivery is not associated with intra-partum MTCT risk.

An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence

Epidemiological studies have demonstrated that co-infections of herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis occur in vivo. Data from a tissue culture model of C. trachomatis/HSV-2 co-infection indicate that viral co-infection stimulates the formation of persistent chlamydiae. Transmission electron microscopic (TEM) analyses demonstrated that in both HeLa and HEC-1B cells, co-infection caused developing chlamydiae to exhibit swollen, aberrantly shaped reticulate bodies (RBs), characteristically observed in persistence. Additionally, HSV-2 co-infection suppressed production of infectious chlamydial elementary bodies (EBs) in both host cell types. Co-infection with HSV type 1 (HSV-1) produced similar morphologic alterations and abrogated infectious EB production. These data indicate that virus-induced chlamydial persistence was neither host cell- nor virus strain-specific. Purification of crude HSV-2 stocks demonstrated that viral particles were required for coinfection-induced chlamydial persistence to occur. Finally, co-infection with either UV-inactivated, replication-incompetent virus or replication-competent HSV-2 in the presence of cyclohexamide reduced chlamydial infectivity without altering chlamydial genomic DNA accumulation. These data demonstrate that productive viral replication is not required for the induction of chlamydial persistence and suggest that HSV attachment and entry can provide the necessary stimulus to alter C. trachomatis development.

HIV/AIDS: Management of Herpes Simplex Virus Type 2 Infection in HIV Type 1–Infected Persons

Human immunodeficiency virus type 1 (HIV-1)-infected persons have high rates of herpes simplex virus type 2 (HSV-2) infection, ranging from 50% to 90% in studies of HIV-infected populations from different parts of the world. Genital herpes in persons with HIV type 1 (HIV-1) infection is associated with more-severe and chronic lesions, as well as increased rates of asymptomatic genital shedding of HSV-2. Nucleoside analogues (acyclovir, valacyclovir, and famciclovir) decrease the frequency and severity of HSV-2 recurrences and asymptomatic HSV-2 reactivation and are effective, safe, well-tolerated drugs in patients with HIV-1 infection. These anti-HSV drugs may result in additional clinical and public health benefits for persons with HIV-1 and HSV-2 coinfection by decreasing HIV-1 levels in the blood and genital tract. Given these benefits, HIV-1-infected persons should be routinely tested for HSV-2 infection using type-specific serologic tests. Persons with HSV-2 infection should be offered HSV-2 education and treatment options. Studies to quantify the potential clinical and public health benefits of treating individuals who have HIV-1 and HSV-2 coinfection with anti-HSV therapy are underway.

Chlamydia trachomatis enters a viable but non-cultivable (persistent) state within herpes simplex virus type 2 (HSV-2) co-infected host cells

Epidemiological and clinical studies have shown that double infection with herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis occurs in vivo. We hypothesized that co-infection would alter replication of these agents. To test this hypothesis, HeLa cells were infected with C. trachomatis serovar E, followed 24 h later by HSV-2 strain 333. Transmission electron microscopic (TEM) analyses indicated that, by 10 h after HSV addition, reticulate bodies (RBs) in co-infected cells were swollen, aberrantly shaped and electron-lucent. In infectious titre assays, HSV-2 co-infection abrogated production of infectious chlamydial progeny. Western blot analyses indicated that accumulation of chlamydial major outer membrane protein (MOMP) was decreased by HSV co-infection while accumulation of chlamydial heat-shock protein 60-1 (HSP60-1) was increased. Polymerase chain reaction (PCR) experiments indicated that chlamydial genome copy number was unaltered by HSV-2 superinfection. Semi-quantitative, reverse transcription PCR (RT-PCR) experiments demonstrated that levels of chlamydial groEL, ftsK, ftsW, dnaA and unprocessed 16S rRNA transcripts were not changed by HSV-2 super-infection. These data indicate that HSV-2 superinfection drives chlamydia into a viable but non-cultivable state, which is the hallmark of persistence. Because chlamydial HSP60-1 has been associated with immunopathology in vivo, these results also suggest that disease severity might be increased in co-infected individuals.

Chronic herpes simplex virus type-2 infection and HIV viral load

By December 2003, the estimated adult HIV/AIDS prevalence rate in sub-Saharan Africa was 7.5-8.5%, and rates of herpes simplex virus type-2 (HSV-2) infection among adults aged >30 years ranged from 60% to 82%. However, little is known about the natural history of HIV/HSV-2 co-infection in this population. We evaluated HIV viral load and CD4+ cell counts among persons with and without chronic HSV-2 co-infection in a cross-sectional study of HIV-infected persons not receiving antiretroviral therapy. HSV-2 and HIV co-infection was associated with a 0.3 log copies/mL higher HIV viral load compared with persons without HSV-2 infection (P=0.014). Chronic HSV-2 infection may have a negative effect on the clinical course of persons with HIV.