Epstein-Barr Virus, But Not Human Papillomavirus, Is Associated With Preinvasive and Invasive Ocular Surface Squamous Neoplasias in Zambian Patients.

Background: The etiopathogenesis of ocular surface squamous neoplasia (OSSN) is not fully understood. We assessed the frequency of oncogenic viruses in OSSN by immunohistochemistry (IHC) and polymerase chain reaction (PCR) for human papillomavirus (HPV), Epstein-Barr virus (EBV), Merkel cell polyomavirus (MCPyV), Kaposi sarcoma virus, and adenovirus. Cases from Zambia were prospectively enrolled using a cross-sectional study design between November 2017 and March 2020. Methods: Demographic and clinical data [age, sex, HIV status, antiretroviral therapy (ART) history, CD4 count, plasma viral load] and tumor biopsies were collected from 243 consenting patients. Tumor samples were bisected, and half was used for DNA isolation, while the other half was formalin fixed and paraffin embedded (FFPE) for histopathology analysis. The expressions of latent EBV nuclear antigen 1 (EBNA1), CDKN2A/p16INK4A (p16), and MCPyV large T-antigen (LT) were tested by IHC. Multiplex PCR was used to detect 16 HPV genotypes and four other DNA tumor viruses [Kaposi's sarcoma-associated herpesvirus (KSHV), EBV, MCPyV, and adenovirus]. Relationships between HIV status, viral DNA and protein expression, and tumor grades were determined by statistical analysis. Results: OSSN tumors from patients were 29.6% preinvasive and 70.4% invasive. Patients presented with unilateral tumors that were 70.4% late stage (T3/T4). OSSN patients were HIV positive (72.8%). IHC on 243 FFPE biopsies resulted in the detection of EBNA1 (EBV), p16 high-risk HPV (HR-HPV), and MCPyV LT expression in 89.0%, 4.9%, and 0.0%, respectively. EBNA1 was expressed in all grades of preinvasive [cornea-conjunctiva intraepithelial neoplasia (CIN)1, 100%; CIN2, 85.7%; CIN3, 95.8%; and carcinoma in situ (CIS), 83.8%] and in invasive (89.2%) OSSN. PCR on 178 samples detected EBV, HR-HPV, and MCPyV in 80.3%, 9.0%, and 13.5% of tumors, respectively. EBV was detected in all grades of preinvasive and invasive OSSN. EBV detection was associated with high HIV viral loads (p = 0.022). HR-HPV was detected in 0.0% CIN1, 0.0% CIN2, 5.6% CIN3, 13.0% CIS, and 7.0% invasive OSSN. Conclusions: Our findings of EBV DNA and EBNA1 protein in all the grades of preinvasive and especially invasive OSSN are consistent with a potential causal role for EBV in OSSN. A role of HPV in OSSN was not clearly established in this study.

Similar Immunological Profiles Between African Endemic and Human Immunodeficiency Virus Type 1-Associated Epidemic Kaposi Sarcoma (KS) Patients Reveal the Primary Role of KS-Associated Herpesvirus in KS Pathogenesis.

BACKGROUND: Kaposi sarcoma (KS)-associated herpesvirus (KSHV) is etiologically linked to all KS forms, but mechanisms underlying KS development are unclear. The incidence of KS in human immunodeficiency virus type 1-infected (HIV-1+) individuals implicates immune dysregulation; however, the lack of characterization of KSHV immune responses in endemic KS makes the role of HIV-1 unclear. The study objective was to investigate the HIV-1 and KSHV roles in viral nucleic acid detection, antibody responses, and cytokine responses in polymerase chain reaction-confirmed epidemic KS and endemic KS patients and non-cancer controls from sub-Saharan Africa. METHODS: KSHV viral DNA (vDNA), total anti-KSHV antibody, KSHV neutralizing antibody (nAb), and cytokines were quantified. RESULTS: KSHV vDNA was detectable in tumors but variably in plasma and peripheral blood mononuclear cells. Consistent with elevated antibody-associated cytokines (interleukin [IL] 6, IL-5, and IL-10), nAb titers were higher in epidemic KS and endemic KS patients than in controls (P < .05). Despite HIV-1 coinfection in epidemic KS, nAb titers were similar between epidemic KS and endemic KS patients (P = 0.3). CONCLUSIONS: Similarities in antibody and cytokine responses between epidemic and endemic KS patients suggest that KSHV drives KS pathogenesis, whereas HIV-1 exacerbates it.

Seroprevalence of human herpesvirus 8 and its impact on the hemoglobin level in patients of end stage of renal diseases.

The incidence of Kaposi's sarcoma (KS) is increasing among renal transplant recipients. Patients with end-stage renal disease (ESRD) are immunocompromised and are candidates for renal transplantation, but HHV8 seroprevalence in ESRD patients has not been well documented. A cross-sectional study of 286 ESRD patients and 281 matched subjects without kidney disease was conducted at the First People's Hospital of Huzhou, Zhejiang province to explore the epidemiologic features of HHV8 among ESRD patients in China. Blood samples were collected and HHV8 antibodies and serologic indices were measured. The seroprevalence of HHV8 was 15.3% for ESRD patients and 8.9% for the comparison group. A significant difference in the geometric mean titer (GMT) of the HHV8 antibodies was detected between ESRD patients and the comparison group (617.1 vs 291.7; P = 0.042). The average level of hemoglobin was 11.56 ± 1.78 g/dL for the ESRD group and 13.73 ± 1.42 g/dL for the comparison group, (P > 0.05). Multiple linear regression revealed a negative association between HHV8 infection and plasma hemoglobin concentration (ß = -0.682, P = 0.036). We found a higher HHV8 prevalence and a higher level of HHV8 antibody GMT in ESRD patients than the comparison group, which indicate a high risk of posttransplantation KS.

Global epidemiology of human herpesvirus 8 in men who have sex with men: A systematic review and meta-analysis.

Men who have sex with men (MSM) were highly vulnerable to HIV/AIDS and Human herpes virus 8 (HHV8), while the epidemiologic features of HHV8 among MSM remain obscure. We therefore performed a systematic review and meta-analysis to assess the burden of HHV8 in MSM. Electronic databases were searched for publications on HHV8 epidemiologic characteristics among MSM. Random-effect meta-analysis was applied to combine the HHV8 seroprevalence in MSM and odds ratios (ORs) for associated risk factors. Meta-regression and stratified analyses were performed to detect the potential sources of heterogeneity. The pooled HHV8 seroprevalence in MSM was 33.0% (95%CI 29.2%-37.1%). Significant factors associated with HHV8 included HIV (OR 3.70, 95%CI 2.93-4.67), STDs (OR 2.32, 95%CI 1.82-2.97), and high risk sexual behaviors (OR 1.50, 95%CI 1.17-1.92). Race (OR 1.44, 95%CI 0.94-2.12) and multiple sexual partners (OR 1.61, 95%CI 0.95-2.72) were also associated with HHV8 (P < 0.10). We found no significant association between IDU and HHV8 (OR 1.44, 95%CI 0.06-32.47). HHV8 is highly prevalent among MSM and the high risk behaviors may facilitate the transmission of this virus. This situation could be of significant public health importance, especially in the context of HIV coinfection.

Association of Household Food- and Drink-Sharing Practices With Human Herpesvirus 8 Seroconversion in a Cohort of Zambian Children.

Background: Human herpesvirus 8 (HHV-8) infection occurs in early childhood and is associated with human immunodeficiency virus type 1 (HIV-1) infection and risk for Kaposi sarcoma, but behaviors associated with HHV-8 transmission are not well described. Methods: We enrolled and followed a prospective cohort of 270 children and their household members to investigate risk factors for HHV-8 transmission in Lusaka, Zambia. Results: We report an incidence of 30.07 seroconversions per 100 child-years. Independent risk factors for HHV-8 incident infection included having a child who shared utensils with a primary caregiver (hazards ratio [HR], 2.33; 95% confidence interval [CI], 1.49-7.14), having an increasing number of HHV-8-infected household members (HR, 1.27; 95% CI, 1.09-2.79), and having ≥5 siblings/children in the household (HR, 2.24; 95% CI, 1.03-4.88). Playing with >5 children a day was protective against infection (HR, 0.54; 95% CI, .33-0.89), as was increasing child age (HR, 0.96; 95% CI, .93-.99). Conclusions: This is the first study to find a temporal association between limited child feeding behaviors and risk for HHV-8 infection. Child food- and drink-sharing behaviors should be included in efforts to minimize HHV-8 transmission, and households with a large number of siblings should receive additional counseling as childhood infections occur in the home context.

Was Kaposi's sarcoma-associated herpesvirus introduced into China via the ancient Silk Road? An evolutionary perspective.

Kaposi's sarcoma-associated herpesvirus (KSHV) has become widely dispersed worldwide since it was first reported in 1994, but the seroprevalence of KSHV varies geographically. KSHV is relatively ubiquitous in Mediterranean areas and the Xinjiang Uygur Autonomous Region, China. The origin of KSHV has long been puzzling. In the present study, we collected and analysed 154 KSHV ORF-K1 sequences obtained from samples originating from Xinjiang, Italy, Greece, Iran and southern Siberia using Bayesian evolutionary analysis in BEAST to test the hypothesis that KSHV was introduced into Xinjiang via the ancient Silk Road. According to the phylogenetic analysis, 72 sequences were subtype A and 82 subtype C, with C2 (n = 56) being the predominant subtype. The times to the most recent common ancestors (tMRCAs) of KSHV were 29,872 years (95% highest probability density [HPD], 26,851-32,760 years) for all analysed sequences and 2037 years (95% HPD, 1843-2229 years) for Xinjiang sequences in particular. The tMRCA of Xinjiang KSHV was exactly matched with the time period of the ancient Silk Road approximately two thousand years ago. This route began in Chang'an, the capital of the Han dynasty of China, and crossed Central Asia, ending in the Roman Empire. The evolution rate of KSHV was slow, with 3.44 × 10-6 substitutions per site per year (95% HPD, 2.26 × 10-6 to 4.71 × 10-6), although 11 codons were discovered to be under positive selection pressure. The geographic distances from Italy to Iran and Xinjiang are more than 4000 and 7000 kilometres, respectively, but no explicit relationship between genetic distance and geographic distance was detected.

Seroprevalence of antibodies against Kaposi's sarcoma-associated herpesvirus among HIV-negative people in China.

BACKGROUND: Little information on the prevalence of Kaposi's sarcoma associated herpesvirus (KSHV) among HIV-negative individuals is available from Asia. METHODS: In the present study, we report findings from a new survey of KSHV in 983 HIV-negative male migrants from Shanghai and their combination with previous similar surveys of 600 female migrants, 600 female sex-workers (FSW), 1336 sexually transmitted infection (STI) clinic male patients, 439 intravenous drug-users (IVDU), and 226 men having sex with men (MSM) from China. KSHV-specific antibodies against latent and lytic antigens were assessed using Sf9 and BC3 monoclonal immunofluorescence assay. Age-adjusted prevalence ratios (PR) and 95% confidence interval (95% CI) for KSHV-positivity were estimated using Poisson regression. RESULTS: In total, 4184 HIV-negative participants were included. KSHV prevalence ranged from 9.8% (95% CI: 7.9%-11.7%) in male migrants to 32.3% (95% CI: 24.1%-34.1%) in MSM. IVDU show intermediate level (17.5%, 95%CI: 14.1%-21.4%). KSHV was associated with syphilis (PR = 2.03, 95% CI: 1.38-2.98) in MSM but not in other groups. An association with human herpes virus 2 was also found among MSM (PR = 1. 83, 95%: 1.22-2.75) but not in migrant workers or FSW. CONCLUSIONS: KSHV prevalence in HIV-negative heterosexuals, FSW, and STI male patients from China is approximately 10%, but 2- and 3-fold higher in IVDU and MSM, respectively. Associations of KSHV with STIs among MSM only suggest that sexual transmission of the virus is important in MSM but not in heterosexuals.

Seroprevalence of Kaposi's sarcoma-associated herpesvirus among HIV-infected Uygurs in Xinjiang, China.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma (KS), which primarily affects human immunodeficiency virus (HIV)-infected adults with advanced immunodeficiency. Xinjiang province in China is an endemic area for Kaposi's sarcoma (KS), however, currently, only limited data for KSHV infection among HIV-infected individuals living in this endemic area is available. A cross-sectional study of 86 HIV positive participants was conducted in Xinjiang, China from 2014 through 2015. Plasma samples were collected and screened for KSHV and HIV infection. HIV pol gene and KSHV ORF-K1 gene were amplified and sequenced, genotypes were determined by phylogenetic analysis. Over all, prevalence was 48.9% (42/86; 95%CI 38.4-59.3%) for KSHV. Only CRF07_BC subtype has been identified among all these HIV positive individuals, while the subtype A and C of KSHV were detected in the participants. Meanwhile, we found that those with high CD4 counts (>500) showed a lower anti-KSHV titer, compared with other groups. Our study indicated a high prevalence of KSHV among HIV positive individuals in Xinjiang, China. Thus, management of HIV/AIDS patients should include KSHV screen and should consider the risk of KSHV-associated malignancies.

High seroprevalence of human herpesvirus 8 and herpes simplex virus 2 infections in men who have sex with men in Shanghai, China.

Men who have sex with men (MSM) are an important risk group for human immunodeficiency virus (HIV) and at high-risk of herpesvirus infection. However, limited information on epidemiologic patterns of HHV8 and HSV2 among MSM is available in mainland China. A cross-sectional study of 486 participants was performed in Shanghai, China from January 2013 to December 2014 to explore epidemiologic features of HHV8 and HSV2 among MSM. Serum samples were collected and tested for HHV8 by immunofluoresence assay and HSV2 antibodies by ELISA. Logistic regression analysis was conducted to determine the risk factors of HHV8 and HSV2 infections, respectively. The overall seroprevalence was 23.0% for HHV8 infection and 22.4% for HSV2 infection. HHV8 seropositivity was significantly associated with being a money boy (odds ratio (OR) = 1.64; 95%CI: 1.04-2.58), having a steady sex partner (OR = 0.52; 95%CI: 0.31-0.87), having ever had anal sex with men (OR = 2.05; 95%CI: 1.09-3.86), and being HSV2 positive (OR = 2.14; 95%CI: 1.22-3.76). HSV2 seropositivity was significantly associate with being positive for HIV (OR = 2.12; 95%CI: 1.01-4.42), syphilis (OR = 1.98; 95%CI: 1.12-3.52), HHV8 (OR = 2.17; 95%CI: 1.24-3.83), and was marginally associated with being a money boy (OR = 1. 61; 95%CI: 0.97-2.86), and having ever had unprotected casual sex (OR = 1. 72; 95%CI: 0.99-2.99). HHV8 and HSV2 infections are common in Chinese MSM. It is important to implement programs for preventing herpes virus infection among MSM, particularly high-risk groups such as money buys. And protected sexual intercourse should be propagated. J. Med. Virol. 89:887-894, 2017. © 2016 Wiley Periodicals, Inc.

Longitudinal analysis of the humoral response to Kaposi's sarcoma-associated herpesvirus after primary infection in children.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent for Kaposi's sarcoma (KS)-one of the most common pediatric cancers in sub-Saharan Africa-however, the factors that lead to disease progression are not fully understood. HIV infection, immunosuppression, and high KSHV viral load increase the risk of developing KS, suggesting that the loss of an effective anti-KSHV immune response may be an important risk factor. However, very little is known about the KSHV-specific immune response prior to KS and less is known about the anti-KSHV immune response during the very early stages of infection. We therefore prospectively followed a cohort of 86 Zambian children for 2 years after primary KSHV seroconversion to characterize the humoral immune response during the early stages of KSHV infection. Plasma, peripheral blood mononuclear cells, and oral swabs were collected from patients every 3 months and analyzed for KSHV-specific antibodies and presence of viral DNA. We observed an approximately 40% KSHV seropositive rate among infected children at time points after primary seroconversion, indicating that seroreversion is common after primary KSHV infection. At the time of primary KSHV seroconversion HIV-infected ART-naïve children had a more robust KSHV antibody response compared to HIV-infected children taking ART and HIV-uninfected children. Conversely, the longitudinal anti-KSHV antibody response was highly variable and did not correlate with available clinical information, HIV/ART status, or presence of KSHV DNA. Collectively, our data suggest that there is limited impact by the variations in the humoral immune response in young children after infection. J. Med. Virol. 88:1973-1981, 2016. © 2016 Wiley Periodicals, Inc.

Prevalence of Kaposi's sarcoma associated herpesvirus among men attending sexually transmitted infections clinics in Anhui, China.

Kaposi's sarcoma-associated herpesvirus (KSHV) may be transmitted via sexual contacts, but limited information is available on KSHV infection status among sexually transmitted infection (STI) patients in China. The main objective of the present study was to determine the KSHV seroprevalence and its risk factors among male STI patients. A cross-sectional survey was conducted in three prefectures of Anhui province, China, between June and September 2013. A total of 1,600 male patients who visited an STI clinic were invited, and 1,372 participated in the study. Data were abstracted from the medical records for all the patients. Blood samples were collected and tested for antibodies to KSHV, HIV, HCV, and syphilis. Factors associated with KSHV seropositivity were examined using multivariable logistic regression analysis. The overall prevalence of KSHV, HIV, HCV, and syphilis was 13.3%, 0.7%, 0.6%, and 12.5%, respectively. After adjusting for potential confounders, KSHV infection was significantly associated with ever having anal sex with men (19 out of 30 males, OR: 8.64, 95%CI: 1.92-38.79) and HIV infection (six out of nine HIV-positive individuals, OR: 8.39, 95%CI: 1.80-39.04). There were no significant associations of KSHV infection with drug use, heterosexual sex behaviors, syphilis, and HCV. Our finding has shown that a relatively moderate prevalence of KSHV was found among male STI patients. While an increased risk for KSHV infection was observed among participants with homosexual contacts. Routine KSHV testing is recommended for male individuals attending STI clinics.

Effects of Antiretroviral Therapy on Kaposi's Sarcoma-Associated Herpesvirus (KSHV) Transmission Among HIV-Infected Zambian Children.

BACKGROUND: The risk of Kaposi's sarcoma-associated herpesvirus (KSHV) acquisition among children is increased by HIV infection. Antiretroviral therapy (ART) was recently made widely available to HIV-infected children in Zambia. However, the impact of early ART on KSHV transmission to HIV-infected children is unknown. METHODS: We enrolled and followed a cohort of 287 HIV-exposed, KSHV-negative children under 12 months of age from Lusaka, Zambia, to identify KSHV seroconversion events. Potential factors associated with KSHV infection-with an emphasis on HIV, ART, and immunological measures-were assessed through structured questionnaires and blood analyses. Incidence rate, Kaplan-Meier, and multivariable Cox regression models were used to assess differences in time to event (KSHV seroconversion) between groups. All statistical tests were two-sided. RESULTS: During follow-up, 151 (52.6%) children underwent KSHV seroconversion. Based on 3552 months of follow-up, we observed similar KSHV incidence rates between HIV-infected and uninfected children. Among HIV-infected children, ART-naïve children had statistically significantly increased risk of KSHV acquisition (adjusted hazard ratio [AHR] = 5.04, 95% confidence interval [CI] = 2.36 to 10.80, P < .001). Time-updated CD4(+) T-cell percentage was also statistically significantly associated with risk of KSHV acquisition (AHR = 0.82, 95% CI = 0.74 to 0.92, P < .001), such that each 5% increase of CD4(+) T-cells represented an 18% decrease in risk of acquiring KSHV. CONCLUSIONS: Our data suggest that early ART and prevention of immune suppression reduce the risk of KSHV acquisition among HIV-infected children in an area where both viruses are highly endemic. This study highlights the importance of programs in Africa to provide children with ART immediately after HIV infection is diagnosed.

Epidemiology and transmission of Kaposi's sarcoma-associated herpesvirus.

This review summarizes the current knowledge pertaining to Kaposi sarcoma-associated herpesvirus (KSHV) epidemiology and transmission. Since the identification of KSHV twenty years ago, it is now known to be associated with Kaposi's sarcoma (KS), primary effusion lymphoma, and multicentric Castleman's disease. Many studies have been conducted to understand its epidemiology and pathogenesis and their results clearly show that the worldwide distribution of KSHV is uneven. Some geographical areas, such as sub-Saharan Africa, the Mediterranean region and the Xinjiang region of China, are endemic areas, but Western Europe and United States have a low prevalence in the general population. This makes it imperative to understand the risk factors associated with acquisition of infection. KSHV can be transmitted via sexual contact and non-sexual routes, such as transfusion of contaminated blood and tissues transplants, or via saliva contact. There is now a general consensus that salivary transmission is the main route of transmission, especially in children residing in endemic areas. Therefore, there is a need to better understand the sources of transmission to young children. Additionally, lack of animal models to study transmission, gold standard serological assay and the lack of emphasis on endemic KS research has hampered the efforts to further delineate KSHV transmission in order to design effective prevention strategies.

Seroprevalence of human herpesvirus 8 and hepatitis C virus among drug users in Shanghai, China.

To elucidate and compare the seroprevalence of human herpesvirus 8 (HHV8) and hepatitis C virus (HCV) among Chinese drug users, a cross-sectional study of 441 participants, was conducted in Shanghai, China, from 2012 through 2013. Seventy-seven (17.5%) participants were found to be positive for HHV8 antibodies, while 271 (61.5%) participants were positive for HCV. No significant association between HHV8 seropositivity and drug use characteristics, sexual behaviors, HCV, or syphilis was observed. In contrast, a statistically significant association between HCV seropositivity and injected drug history (OR, 2.18, 95% CI 1.41-3.37) was detected, whereas no statistically significant association between HCV seropositivity and syphilis infection (OR, 7.56, 95% CI 0.94-60.57) were observed. Pairwise comparisons showed no significant differences between latent and lytic antibodies regarding HCV and HHV8 serostatus. The study demonstrated a moderate but elevated prevalence of HHV8 infection among drug users. The discordance between HHV8 and HCV infections suggests that blood borne transmission of HHV8 might not be the predominant mode of transmission in this population, which is in contrast to HCV.

Kaposi`s sarcoma associated herpesvirus infection among female sex workers and general population women in Shanghai, China: a cross-sectional study.

BACKGROUND: Limited information on epidemiologic patterns of KSHV, with none focusing on heterosexual transmission, is available in mainland China. To clarify this, a cross-sectional study was conducted among a group of female sex workers (FSW) and general population women (GW) in Shanghai, China. METHODS: An anonymous questionnaire interview was administrated among 600 FSW and 600 GW. Blood samples were collected and tested for antibodies to KSHV, HSV-2, HIV, syphilis and HBsAg. Correlates of KSHV and HSV-2 were examined using multiple logistic regression analysis. RESULTS: None of the study participants were tested positive for HIV. The seroprevalence of KSHV, HSV-2 , HBV and syphilis was 10.0%, 52.2%, 12.3% and 10.5%, respectively for FSW, and was 11.0%, 15.3%, 9.8% and 2.8%, respectively for GW. KSHV seropositivity was not associated with syphilis and HSV-2 infection as well as sexual practices among either FSW or GW. Nevertheless, HSV-2 infection among FSW was independently associated with being ever married (OR = 1.59; 95%CI: 1.04-2.45), >5 years of prostitution (OR = 2.06; 95%CI: 1.16-3.68) and being syphilis positive (OR = 2.65; 95%CI: 1.43-4.93). HSV-2 infection among GW was independently associated with an age of >35 years (OR = 2.29; 95%CI: 1.07-4.93), having had more than 2 sex partners in the prior 12 months (OR = 6.44; 95%CI: 1.67-24.93) and being syphilis positive (OR = 3.94; 95%CI: 1.38-11.23). A gradual increase of prevalence with the prostitution time group was also detected for HSV-2 and syphilis, but not for KSHV. CONCLUSIONS: KSHV is moderately and equivalently prevalent among FSW and GW. Heterosexual contact is not a predominant route for KSHV transmission among Chinese women.

Inactivation and viral load quantitation of human immunodeficiency virus in blood collected into Cyto-Chex(®) BCT blood collection device.

A blood collection tube (Cyto-Chex(®) BCT), which can stabilize white blood cells and immunogenic markers in blood samples, was investigated for its ability to inactivate human immunodeficiency virus (HIV) and stabilize HIV for viral load quantitation. Laboratory-adapted HIV strains were either treated or untreated with the stabilizing reagent present in Cyto-Chex(®) BCT. A dilution of the reagent used to treat virus was 1:66, which was similar to the reagent concentration in Cyto-Chex(®) BCT device when blood was drawn into it. In another experiment, blood was drawn from HIV patients into one acid citrate dextrose (ACD) tube and one Cyto-Chex(®) BCT. At indicated time points, aliquots were taken of treated and untreated viral dilutions and from plasma of HIV-positive patient blood samples and analyzed using reverse transcriptase and TZM-bl cell assays to determine HIV inactivation. In laboratory-adapted HIV strains and HIV-positive patient plasma, HIV was completely inactivated within 2 and 3h of contact with a 1:66 dilution of Cyto-Chex reagent, respectively. Samples from HIV-positive patient plasma showed that viral load was stable in Cyto-Chex(®) BCT for 7 days at room temperature. Therefore, it is concluded that the chemical reagent present in the Cyto-Chex(®) BCT blood collection device is capable of complete inhibition of HIV infectivity in blood samples within 3h and stabilizing the viral load for 7 days at room temperature.

Risk factors for early childhood infection of human herpesvirus-8 in Zambian children: the role of early childhood feeding practices.

BACKGROUND: Human herpesvirus-8 (HHV-8) infection in early childhood is common throughout sub-Saharan Africa with prevalence increasing throughout childhood. Specific routes of transmission have not been clearly delineated, though HHV-8 is present in high concentrations in saliva. METHODS: To understand the horizontal transmission of HHV-8 within households to children, we enrolled for cross-sectional analysis, 251 households including 254 children, age two and under, in Lusaka, Zambia. For all children, plasma was screened for HHV-8 and HIV type I (HIV-1) and health and behavioral questionnaires were completed. Multilevel logistic regression analysis was conducted to assess independent factors for HHV-8 infection in children. RESULTS: Risk factors for HHV-8 infection included increasing number of HHV-8-positive household members [OR = 2.5; 95% confidence interval (CI), 1.9-3.3; P < 0.01] and having a primary caregiver who tested the temperature of food with their tongue before feeding the child (OR = 2.4; 95% CI, 1.93-3.30; P = 0.01). Breastfeeding was protective against infection with HHV-8 for children (OR = 0.3; 95% CI, 0.16-0.72; P < 0.01). CONCLUSIONS: These results indicate that exposure to HHV-8 in the household increases risk for early childhood infection, with specific feeding behaviors likely playing a role in transmission. IMPACT: Interventions to protect children from infection should emphasize the possibility of infection through sharing of foods.

Higher levels of neutralizing antibodies against KSHV in KS patients compared to asymptomatic individuals from Zambia.

Kaposi sarcoma-associated herpesvirus (KSHV) is the etiologic agent for Kaposi Sarcoma (KS), the most common cancer diagnosed in HIV- infected patients. The role of neutralizing antibodies in KS pathogenesis and in KSHV infected individuals is not clearly understood. The goal of this study was to investigate and compare the prevalence and titers of neutralizing antibodies in plasma samples from KS patients and KSHV infected asymptomatic individuals from Zambia, a KS endemic region in sub-Saharan Africa. Plasma samples (N = 267) consisting of KS patients (group 1) and asymptomatic individuals (group 2) were collected from Lusaka, Zambia. A flow cytometry based quantitative neutralization assay utilizing recombinant KSHV expressing GFP was used to detect KSHV neutralizing antibodies. Our results show that the overall prevalence of neutralizing antibodies in KS patients (group 1) was 66.7% which was significantly higher than the prevalence of 6.5% present in KSHV infected asymptomatic individuals (group 2). Total antibody titers as well as neutralizing antibodies titers were found to be significantly higher among KS patients. It is likely that higher neutralizing antibodies prevalence and titers in KS patients result from higher levels of antigenic stimulation over time. This study is first to compare prevalence and titers of neutralizing antibodies in participants with and without disease from a KSHV endemic region.