Protein Expression of TLR2, TLR4, and TLR9 on Monocytes in TB, HIV, and TB/HIV.

Toll-like receptors (TLRs) have a critical role in recognizing pathogenic patterns and initiating immune responses against TB and HIV. Previously, studies described the gene expression of TLRs in patients with TB and HIV. Here, we demonstrated TLRs protein expressions and their association with clinical status and plasma markers in TB, HIV, and TB/HIV coinfection. The phenotyping of TLR2, TLR4, and TLR9 on CD14+ monocytes and their subsets were determined by multicolor flow cytometry. Host plasma biomarkers and microbial indices were measured using Luminex Multiplex assay and standard of care tools, respectively. TLR2 expression significantly enhanced in TB, slightly increased in HIV but slightly reduced in TB/HIV coinfection compared to apparently health controls (HC). On the other hand, TLR4 expression was significantly increased in TB, HIV, and TB/HIV compared to HC. Expression of TLR4 was equally enhanced on classical and intermediate monocytes while higher TLR2 expression on intermediate than classical monocytes. TLR4 had a positive correlation pattern with plasma biomarkers while TLR2 had an inverse correlation pattern. TLR4 is associated with disease severity while TLR2 is with the immune-competent status of patients. Our findings demonstrated that the pattern of TLR expression is disease as well as monocyte subset specific and distinct factors drive these differences.

Differential expression of chemokine receptors on monocytes in TB and HIV S.

In the present study, we defined multiple chemokine receptors expressed by classical, intermediate and non-classical monocyte subsets in TB, HIV and TB/HIV co-infection and associate it with the perturbation of monocyte subsets due to the diseases. Peripheral blood mononuclear cells from TB+ (n = 34), HIV+ (n = 35), TB + HIV+ (n = 12), as well as TB-HIV- healthy controls (n = 39), were tested for monocyte phenotyping by flow cytometry. Frequencies of intermediate and non-classical monocytes were significantly higher in TB and/or HIV disease relative to healthy controls. CCR2 and CX3CR1 were significantly higher on monocytes in TB disease, whereas CCR4 and CCR5 were present at higher levels in HIV disease. TB/HIV co-infected patients exhibited CCR2, CCR5 and CX3CR1 levels intermediate to TB and HIV subjects, while CCR4 was at a higher level than HIV. Despite the increase in the expression of chemokine receptors due to disease conditions, chemokine receptors maintained their original expression pattern on monocyte subsets. Our data provided new insight into the disease-specific but not monocyte subsets-specific modulation of chemokine receptors in TB and HIV.

Phenotypic characterization of Peripheral B cells in Mycobacterium tuberculosis infection and disease in Addis Ababa, Ethiopia.

BACKGROUND: Mortality and morbidity from tuberculosis (TB) remain one of the most important public health issues. Although cell-mediated immunity is the main immune response against Mycobacterium tuberculosis (MTB), the role of B-cells during MTB infection and disease is unclear. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from treatment naïve Pulmonary TB patients (TB, n = 16), latent TB-infected participants (LTBI, n = 17), and healthy controls (HC, n = 19). PBMCs were stained with various fluorescently labeled antibodies to define B-cell subsets using multicolor flow cytometry. RESULTS: Atypical memory B cells (CD19+CD27-CD21-) and circulating marginal zone B-cells (CD19+CD27+CD21+IgM+IgD+CD23-) were significantly higher in active TB when compared to LTBI and HC. CD5+ regulatory B cells (Breg, CD19+CD24hiCD38hiCD5+) and resting B-cells (CD19+CD27+CD21+) in Active TB patients were significantly lower compared to HC and LTBI. Overall, there were no differences in B cell percentages (CD19+), naïve B cells (CD19+CD27-CD21+), Breg (CD19+CD24hiCD38hi), and activated memory B cells (CD19+CD27+CD21-) among the three study groups. CONCLUSIONS: These results indicated that multiple subsets of B cells were associated with TB infection and disease. It will be useful to examine these cell populations for their potential use as biomarkers for TB disease and LTBI.

PDL1 expression on monocytes is associated with plasma cytokines in Tuberculosis and HIV.

INTRODUCTION: PDL1 and its interaction with PD1 is implicated in immune dysfunction in TB and HIV. The expression of PDL1 on multiple subsets of monocytes as well as their associations with cytokines and microbial products have not been well studied. METHOD: HIV (TB-HIV+), TB (TB+HIV-) and TB/HIV co-infected (TB+HIV+) patients as well as apparently healthy controls (TB-HIV-) were recruited. TB and HIV patients were treatment naïve while TB/HIV patients were both ART naïve and experienced but not yet started TB therapy. Monocyte subsets were evaluated for PDL1 expression by flow cytometry; plasma TNFα, IL6, IP10, IFNγ and IL10 were measured by Luminex; and cytokine mRNA from purified monocytes quantitated by qPCR. The association of PDL1 with cytokines, clinical and microbial indices, including HIV viral load, TB smear microscopy and TB urinary lipoarabinomannan (LAM) were assessed. RESULTS: Monocyte expression of PDL1 was significantly higher in TB, HIV and TB/HIV co-infected patients compared with healthy controls (p = 0.0001), with the highest levels in TB/HIV co-infected patients. The highest expression of PDL1 was on intermediate (CD14+CD16+) monocytes in all participant groups. PDL1 strongly correlated with HIV viral load in TB/HIV while weakly correlated in HIV. PDL1 levels moderately correlated with plasma TNFα, IL6, IP10, IFNγ and IL10 level in TB subjects whereas weakly correlated with TNFα and IP10 in HIV patients. However, cytokine mRNA from purified monocytes showed no association with either plasma cytokines or monocyte PDL1 expression, implying that if cytokines modulate PDL1, they are likely not originating from circulating monocytes themselves. These results underscore the importance of further characterization of multiple monocyte subsets and their phenotypic and functional differences in different disease states.

Human herpesvirus 8 infection in HIV-1-infected and uninfected pregnant women in Ethiopia.

BACKGROUND: Human Herpes Virus (HHV-8) is related to Kaposi Saracoma, an opportunistic infection occurring with HIV infection. Little is known about the seroepidemiology of Human Herpesvirus 8 (HHV-8) infection among Ethiopian women, even though women are a major HIV risk group in Ethiopia. OBJECTIVES: This study aimed at determining the seroprevalence of HHV-8 infection in HIV-1-infected and uninfected pregnant women in five selected regions of Ethiopia. METHODS: A cross-sectional study was conducted from December 2006 to June 2007 where pregnant women were recruited after age-matching in groups. A total of 400 pregnant women were enrolled, with 200 being HIV-infected and 200 being HIV-uninfected Sera were screened for IgG lytic antibody to HHV-8 using an Indirect Fluorescence Assay (IFA) in Virology Unit of Ethiopian Health and Nutrition Research Institute (EHNR1). RESULTS: Of 400 pregnant women attending antenatal clinic (ANC) testing sites of five regions in Ethiopia, 212 (53.0%) were positive for HHV-8 IgG lytic antibody. There was a high prevalence of HHV-8 infection among HIV-1-infected pregnant women (138, 69.0%) as compared with HIV-1-uninfected pregnant women (74, 37.0%). CONCLUSION: The study shows a high prevalence of HHV-8 infection among HIV-1-infected pregnant women as compared with HIV-1-uninfected pregnant women. Therefore, creating awareness and educating women on safe sexual practice and avoiding deep kissing may be a fundamental ways to limit the roots of transmission. Moreover, initiating strong antiretroviral therapy (ART) for HIV infected women would be best treatment prior to the development of Kaposi's sarcoma (KS).

RNA detection and subtype C assessment of HIV-1 in infants with diarrhea in Ethiopia.

In the absence of chemoprophylaxis, HIV-1 transmission occurs in 13-42% of infants born to HIV-1 positive mothers. All exposed infants acquire maternal HIV-1 antibodies that persist for up to 15 months, thereby hampering diagnosis. In resource limited settings, clinical symptoms are the indices of established infection against validated laboratory-based markers. Here we enrolled 1200 children hospitalized for diarrheal and other illnesses. 20-25% of those tested, aged 15 months or younger, were found to be HIV-1-seropositive. Where sufficient plasma was available, HIV-1 RNA detection was performed using a subtype-insensitive assay, with 71.1% of seropositive infants presenting with diarrhea showing positive. From sub-typing analysis, we identified that viruses of the C' sub-cluster were predominated amongst infants. Although this study may overestimate the HIV-1 frequency through testing symptomatic infants, diarrhea can be seen as a useful marker indicating HIV-1 infection in infants less than 15 months old.