Microbial cancer immunotherapy reprograms hematopoietic stem cells to enhance anti-tumor immunity.

Mycobacterium bovis BCG is the vaccine against tuberculosis and an immunotherapy for bladder cancer. When administered intravenously, BCG reprograms bone marrow hematopoietic stem and progenitor cells (HSPCs), leading to heterologous protection against infections. Whether HSPC-reprogramming contributes to the anti-tumor effects of BCG administered into the bladder is unknown. We demonstrate that BCG administered in the bladder in both mice and humans reprograms HSPCs to amplify myelopoiesis and functionally enhance myeloid cell antigen presentation pathways. Reconstitution of naive mice with HSPCs from bladder BCG-treated mice enhances anti-tumor immunity and tumor control, increases intratumoral dendritic cell infiltration, and synergizes with checkpoint blockade. We conclude that bladder BCG acts systemically, reprogramming HSPC-encoded innate immunity, highlighting the broad potential of modulating HSPC phenotypes to improve tumor immunity. One Sentence Summary: BCG administered in the bladder reprograms bone marrow HSPCs and contributes to tumor control via enhanced myeloid cells.

Chalkophore mediated respiratory oxidase flexibility controls M. tuberculosis virulence.

Oxidative phosphorylation has emerged as a critical therapeutic vulnerability of M. tuberculosis, but it is unknown how M. tuberculosis and other pathogens maintain respiration during infection. M. tuberculosis synthesizes diisonitrile lipopeptide chalkophores that chelate copper tightly, but their role in host-pathogen interactions is also unknown. We demonstrate that M. tuberculosis chalkophores maintain the function of the heme-copper bcc:aa3 respiratory oxidase under copper limitation. Chalkophore deficient M. tuberculosis cannot survive, respire to oxygen, or produce ATP under copper deprivation in culture. M. tuberculosis lacking chalkophore biosynthesis is attenuated in mice, a phenotype that is severely exacerbated by loss of the CytBD alternative respiratory oxidase (encoded by cydAB), revealing a multilayered flexibility of the respiratory chain that maintains oxidative phosphorylation during infection. Taken together, these data demonstrate that chalkophores counter host inflicted copper deprivation and highlight that protection of cellular respiration is a critical virulence function in M. tuberculosis.

Commensal antimicrobial resistance mediates microbiome resilience to antibiotic disruption.

Despite their therapeutic benefits, antibiotics exert collateral damage on the microbiome and promote antimicrobial resistance. However, the mechanisms governing microbiome recovery from antibiotics are poorly understood. Treatment of Mycobacterium tuberculosis, the world's most common infection, represents the longest antimicrobial exposure in humans. Here, we investigate gut microbiome dynamics over 20 months of multidrug-resistant tuberculosis (TB) and 6 months of drug-sensitive TB treatment in humans. We find that gut microbiome dynamics and TB clearance are shared predictive cofactors of the resolution of TB-driven inflammation. The initial severe taxonomic and functional microbiome disruption, pathobiont domination, and enhancement of antibiotic resistance that initially accompanied long-term antibiotics were countered by later recovery of commensals. This resilience was driven by the competing evolution of antimicrobial resistance mutations in pathobionts and commensals, with commensal strains with resistance mutations reestablishing dominance. Fecal-microbiota transplantation of the antibiotic-resistant commensal microbiome in mice recapitulated resistance to further antibiotic disruption. These findings demonstrate that antimicrobial resistance mutations in commensals can have paradoxically beneficial effects by promoting microbiome resilience to antimicrobials and identify microbiome dynamics as a predictor of disease resolution in antibiotic therapy of a chronic infection.

Experiences and training needs of healthcare providers involved in the care of Ghanaian adolescents living with HIV: an interventional study.

BACKGROUND: Caring for adolescents living with HIV/AIDS (ALHIV) can be overwhelming due to their unique needs. Ghana is currently among nine countries in West and Central Africa contributing to 90% of new paediatric infections in the sub-region with a growing population of ALHIV. Regardless, gaps in paediatric related care including healthcare providers (HCPs) capacity issues have been identified. This study sought to assess the competencies of adolescent-oriented healthcare providers before, and after interventionist training to inform recommendation that would guide the psychosocial care they give to ALHIV. METHODS: The study adopted a mixed methods approach with a non-randomized interventional study involving three-phase multi-methods. The sample consisted of 28 adolescent-oriented and multi-disciplinary healthcare providers at the Cape Coast Teaching Hospital (CCTH) in Ghana. Data were obtained in three phases, namely, a baseline survey, interventionist training, and post-training in-depth interviews. Quantitative data were analyzed using Stata version 13 for descriptive analysis while the qualitative data were analyzed thematically using NVivo version 11. RESULTS: Although the majority of the HCPs claim to be knowledgeable about adolescent health issues (n = 21, 75.0%), only about a third (n = 10, 35.7%) could correctly define who an adolescent is. The majority (n = 18, 64.3%) had not received any training on how to work with the adolescent client. The main areas identified for improvement in the ALHIV care in phase 1 included issues with psychosocial assessment, communication and treatment adherence strategies, creating an adolescent-friendly work environment, and availability of job aids/protocols. During the post-training interviews, participants reported an improved understanding of the characteristics of an adolescent-friendly site and basic principles for ALHIV care. They were also able to correctly describe the widely used adolescent health assessment tool; the HEEADSSS. Post intervention interviews also revealed HCPs perception on increased practice related confidence levels and readiness to implement new knowledge and skills gained. CONCLUSION: This study has shown that targeted training on routine ALHIV care is effective in increasing HCPs knowledge, skills and confidence. Addressing the healthcare system/facility related gaps serves as an impetus for improved ALHIV care among HCPs.

Knowledge and experience of a cohort of HIV-positive and HIV-negative Ghanaian women after undergoing human papillomavirus and cervical cancer screening.

BACKGROUND: Cervical cancer is the most common cancer in women in Ghana, but knowledge and experience of women who have had cervical screening is under-evaluated. This study examined knowledge and understanding of HPV and cervical cancer and evaluated experiences of screening in a cohort of women of mixed HIV status. METHODS: This was a mixed methods study using questionnaires and focus group discussions, with a knowledge score constructed from the questionnaire. HIV-positive and HIV-negative women were recruited from a larger cervical screening study in Ghana and were interviewed 6 months after receiving screening. Quantitative data was analyzed and triangulated with qualitative data following thematic analysis using the framework approach. RESULTS: A total of 131 women were included (HIV-positive, n = 60). Over 80% of participants had a knowledge score deemed adequate. There was no difference between HIV-status groups in overall knowledge scores (p = 0.1), but variation was seen in individual knowledge items. HIV-positive women were more likely to correctly identify HPV as being sexually-transmitted (p = 0.05), and HIV negative women to correctly identify the stages in developing cervical cancer (p = < 0.0001). HIV-positive women mostly described acquisition of HPV in stigmatising terms. The early asymptomatic phase of cervical cancer made it difficult for women to define "what" cancer was versus "what" HPV infection was. All women expressed that they found it difficult waiting for their screening results but that receiving information and counselling from health workers alleviated anxiety. CONCLUSIONS: Knowledge of women who had participated in a cervical screening study was good, but specific misconceptions existed. HIV-positive women had similar levels of knowledge to HIV-negative, but different misconceptions. Women expressed generally positive views about screening, but did experience distress. A standardized education tool explaining cervical screening and relevance specifically of HPV-DNA results in Ghana should be developed, taking into consideration the different needs of HIV-positive women.

ART Regimen and Other Sociodemographics Do Not Affect Cytokine Expression in HIV Patients in Ghana.

BACKGROUND: HIV infection is marked by the production of cytokines by infected cells and cells of the immune system. Variations in the levels of cytokine in HIV-infected individuals significantly impact the role of the immune system with the possibility to affect the course of HIV disease by either exacerbating or suppressing HIV replication. AIM: The study sought to investigate the effect of sociodemographic indices, clinical laboratory parameters, and ART regimen on Th1, Th2, and Th17 cytokines in HIV patients. MATERIALS AND METHODS: A total of two hundred (200) HIV patients on either the first or second line of ART were recruited into the study. Sociodemographic indices were collected using researcher-administered questionnaires. Serum concentrations of two major immune-promoting cytokines, IL-12 and IFN-γ, and immune-suppressive cytokines, IL-10 and IL-17, were measured using enzyme-linked immunosorbent assay (ELISA). T-test and chi-square were used to compare mean scores, while correlation (Pearson's correlation) and linear regression analyses were also performed with the statistical significance set at p < 0.05. RESULTS: The mean age of the participants was (45.54 ± 0.7846) years with a greater proportion (84.5%) between 31 and 60 years. The mean interferon-gamma (INF-γ), interleukin- (IL-) 10, interleukin-12, and interleukin-17 were estimated to be 349.9 ± 8.391 pg/ml, 19.32 ± 0.4593 pg/ml, 19.23 ± 0.3960 pg/ml, and 24.6 ± 0.6207 pg/ml, respectively. Although INF-γ and IL-17 levels were relatively higher in males compared to females, it was vice versa for IL-10 and IL-12. However, none of these was statistically significant. Again, no significant difference was observed among all the cytokines stratified by the duration of ART, stage of HIV, and smoking status. Most importantly, stratification by either first- or second-line ART regimens recorded no significant difference in cytokine levels. Age significantly correlated inversely with IFN-γ (r = -0.27, p ≤ 0.001), IL-10 (r = -0.24, p ≤ 0.001), and IL-12 (r = -0.18, p=0.01) while duration on ART significantly correlated inversely with IFN-γ (r = -0.16, p=0.02). CD4 counts at 6 months and 12 months on ART correlated inversely with IL-17 (r = -0.17, p=0.02) and plasma viral load at 1 year (r = -0.22, p ≤ 0.001), respectively. A positive correlation was observed between IFN-γ and IL-12 (r = -0.84, p ≤ 0.001) and IL-17 (r = -0.50, p ≤ 0.001). This positive trend was repeated between IL-10 and IL-12 (r = -0.92, p ≤ 0.001) and IL-17 (r = -0.61, p ≤ 0.001). CONCLUSION: The levels of IFN-γ, IL-12, IL-17, and IL-10 are not significantly affected by sociodemographics and ART regimen. This observation shows that no significant difference was observed in cytokine levels stratified by ART regiments. This means that both regimens are effective in the suppression of disease progression.

Post Hepatitis B vaccination sero-conversion among health care workers in the Cape Coast Metropolis of Ghana.

BACKGROUND: HBV vaccine is known to offer protection against transmission of HBV infection. Health care workers are mandated to have this vaccination as part of their occupational health safety measures. Post vaccination response data for HCWs in our setting is not available. This study therefore aimed to evaluate the anti-HBs titre levels after Hepatitis B vaccination among HCWs from selected heath facilities in the Cape Coast Metropolis, Ghana. METHODS: A multicenter (3 selected sites) analytical cross-sectional study involving 711 HCWs was conducted. Five (5mls) of blood samples were collected from each study participant and the serum used for HBV immunological profile testing anti-HBs quantification by ELISA test (Fortress Diagnostics Limited, Northern Ireland, United Kingdom). Data analyses were performed using Stata version 14.0 software (STATA Corp, Texas USA). RESULTS: The median age of participants was 29 years (IQR = 26-35 years). Majority (80.9%, n = 575) took their vaccination from Government health facilities compared with 19.1% (n = 136) from private vaccination sources. A total of 7 (3 males and 4 females) were found to be HBsAg positive giving prevalence of 1%. In all, 8.2% (n = 58) of the HCWs had anti-HBs titre levels <10IU/ml giving a sero-protection rate of 91.8%. HCWs who received 3 doses of HBV vaccine were more likely to be sero-protected as compared to those who received only one dose in multivariate analysis (aOR = 3.39, 95%CI: 1.08-10.67), p<0.037). Gender, cigarette smoking and alcohol consumption were not found to be associated with sero-protection. CONCLUSION: There is a high HBV vaccine efficacy among HCWs in the Cape Coast Metropolis of Ghana with higher prevalence of anti-HBs titre level associated with full vaccine dose adherence. Post vaccination antibody titre determination could be an integral part of HBV vaccination protocol for HCWs in Ghana.

HIV diagnosis disclosure to infected children and adolescents; challenges of family caregivers in the Central Region of Ghana.

BACKGROUND: Disclosure of Human Immunodeficiency Virus (HIV) to infected older children and adolescents is essential for both personal health maintenance and HIV prevention within the larger population. Non-disclosure of HIV status has been identified as one of the potential barriers to optimum adherence especially in children and adolescents. Like many other countries in the SSA region, Ghana has significant number of children and adolescents infected by HIV, who have increased survival times, due to increased access to ART. However, both family caregivers and healthcare workers face an array of challenges with the disclosure process, including the timing, what information about the child's HIV status should be shared with him/her and how to go about it. The aim of the study was to identify family caregiver factors associated with non-disclosure of HIV status to infected children and adolescents accessing Antiretroviral Therapy (ART) at the three main ART sites within the Central Region of Ghana. METHODS: A quantitative analytical survey was conducted among 103 family caregivers of HIV infected children (aged 6-17 years) assessing ART services in the Central Region of Ghana. Data were analyzed using SSPS version 21. RESULTS: The age range of caregivers was 20-69 years. The study found a low disclosure rate (23.3%) among caregivers. Majority of the caregivers (80.6%) lacked knowledge on the process of disclosure (how and what to tell child), and majority (64%) also had never received guidance about the disclosure process from their healthcare providers. The main barriers to disclosure were caregiver lack of knowledge regarding the disclosure process and when to disclose, the fear of child's reaction, and fear of stigmatization and associated negative social consequences. CONCLUSION: These findings suggest a lesser involvement of health care providers in preparing caregivers for the disclosure process. This therefore highlight the need for the National HIV/AIDS/STI Control Program to strengthen the involvement and training of healthcare providers in HIV diagnosis disclosure to infected children, based on context-specific policy guidelines informed by the WHO recommendations.

Immunologic and virological response to ART among HIV infected individuals at a tertiary hospital in Ghana.

BACKGROUND: The need to study the outcome of Antiretroviral Therapy (ART) among Human Immunodeficiency Virus (HIV) infected individuals in Ghana, a sub-Saharan African country crucial in the era of the "Treat All" policy. The aim of this study was to analyze selected determinants of immunological and virological response to ART among HIV infected individuals in a tertiary facility in Cape Coast, Ghana. METHODS: An analytical cross sectional study with a retrospective component was conducted in the Cape Coast Teaching Hospital (CCTH), Central Region. Clients aged 18 years and above attending the HIV Clinics for ART and who were on ART for 6 months or more were recruited. The viral loads, CD4 count and other socio-demographic data were analyzed using STATA version 13 (STATA Corp, Texas USA). Descriptive analysis was done and presented with appropriate measures of central tendencies. In addition, bivariate and multivariate analysis was carried out with p value of 0.05 interpreted as evidence of association between variables. RESULTS: A total of 440 participants were included in this study with a mean age of 45.5 (±11.6) years. The mean CD4 count at baseline, 6 months on ART and currently at study recruitment were 215.1 cells/mm3 (±152.6), 386.6 cells/mm3 (±178.5), and 579.6 cells/mm3 (±203.0) respectively. After 6 months and 12 months on ART, the number who had achieved viral copies < 1000/ml were 149 (47.0%) and 368 (89.6%) respectively. There was strong evidence of an association between having CD4 count < 350 cells/mm3 after 6 months on ART and having a diagnosis of tuberculosis since HIV diagnosis (aOR 8.5, 95% CI 1.1-73.0, p = 0.05) and clients having plasma viral load > 1000 copies/ml after 6 months on ART (aOR 2.0, 95% CI 1.2-3.2, p = 0.01). CONCLUSION: There was good response to ART among clients, high virological suppression and immunological recovery hence low rates of change to second line ART regimen in this cohort studied. With strict adherence to the national policy on HIV testing, management of positive clients and full implementation of the "Treat All" policy, Ghana could achieve, if nothing at all, the third "90, 90, 90" target by 2020.

Self-collected vaginal sampling for the detection of genital human papillomavirus (HPV) using careHPV among Ghanaian women.

BACKGROUND: Detection of genital HPV DNA is recommended as an important strategy for modern cervical cancer screening. Challenges include access to services, the reliance on cervical samples taken by clinicians, and patient's preference regarding provider gender. The objective of this research was to determine the acceptability, feasibility and performance of alternative self-collected vaginal samples for HPV detection among Ghanaian women. METHODS: A comparative frequency-matched study was conducted in a systematic (1:5) sample of women attending HIV and outpatient clinics in the Cape Coast Teaching Hospital, Ghana. Participants were instructed on self-collection (SC) of vaginal samples using the careHPV brush and a clinician-collected (CC) cervical sample was obtained using a similar brush. Paired specimens were tested for HPV DNA (14 high-risk types) by careHPV assay (Qiagen) and by HPV genotyping (Anyplex II, Seegene). RESULTS: Overall, 194 women of mean age 44.1 years (SD ± 11.3) were enrolled and 191 paired SC and CC results were analysed. The overall HPV detection concordance was 94.2% (95%CI: 89.9-97.1), Kappa value of 0.88 (p < 0.0001), showing excellent agreement. This agreement was similar between HIV positive (93.8%) and negative (94.7%) women. Sensitivity and specificity of SC compared to CC were 92.6% (95%CI: 85.3-97.0) and 95.9% (95%CI: 89.8-98.8) respectively. The highest sensitivity was among HIV positive women (95.7%, 95%CI: 88.0-99.1) and highest specificity among HIV negative women (98.6%, 95%CI: 92.4-100). Overall, 76.3% women found SC very easy/easy to obtain, 57.7% preferred SC to CC and 61.9% felt SC would increase their likelihood to access cervical cancer screening. CONCLUSIONS: The feasibility, acceptability and performance of SC using careHPV support the use of this alternative form of HPV screening among Ghanaian women. This could be a potential new affordable strategy to improve uptake of the national cervical cancer screening program.