Does in utero HIV exposure and the early nutritional environment influence infant development and immune outcomes? Findings from a pilot study in Pretoria, South Africa

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dc.contributor.author White, Marina
dc.contributor.author Feucht, Ute Dagmar
dc.contributor.author Duffley, Eleanor
dc.contributor.author Molokoane, Felicia
dc.contributor.author Durandt, Chrisna
dc.contributor.author Cassol, Edana
dc.contributor.author Rossouw, Theresa M.
dc.contributor.author Connor, Kristin L.
dc.date.accessioned 2021-07-27T06:17:33Z
dc.date.available 2021-07-27T06:17:33Z
dc.date.issued 2020-12-11
dc.description Additional file 1: Supplementary Figure S1. Sequential gating approach for the measurement of CCR2 expression by CD14+ monocytes. The sequential gating approached used was as follows: First, the viable (7-AAD negative; region ‘Viable”) cells were identified using a 7-AAD vs SS Log density plot. A “Viable” region was created around the 7-AAD negative cells. Gated on the “Viable”cells, a SSLog vs FS plot was used to capture intact cells in the “E” region. CD14+ monocytes were identified (“CD14+” region) using a CD14 vs SS Log density plot that were gated on viable, intact cells (“E” region). CD14+ monocytes that express CCR2 were quantified using a CD192 (CCR2) vs SS Log plot. The proportion of CD14+/CCR2+ cells were captured in the “CD14+ CCR2+” region. The gating strategy followed to quantify CCR2 expression by CD16+ neutrophils was similar to what was described for CD14+ monocytes, but instead of identifying CD14+ monocytes, CD16+ neutrophils were identified (“CD16+” region) using a CD16 vs SS Log density plot that were gated on viable, intact cells (“E” region). CD16+ neutrophils that express CCR2 were quantified using a CD192 (CCR2) vs SS Log plot. The proportion of CD14+/CCR2+ cells was captured in the “CD16+ CCR2+” region. en_ZA
dc.description Additional file 2: Supplementary Figure S2. Sequential gating approach for the measurement of CCR2 expression by monocyte subpopulations. Doublets and debris were removed (Region ‘K’) using a FS Area vs FS Height density plot. A 7-AAD vs SS Log density plot, gated on ‘K’ was used to exclude all non-viable cells. Viable cells were captured in region ‘Viable’. Viable CD14+ monocytes were identified (Region ‘CD14+ Monocytes’) using a CD14 APC vs SS Log density plot. Monocyte sub-populations were identified using a CD16 FITC vs CD14 PE density plot gated on viable, CD14+ monocytes. Four monocyte sub-populations were identified: CD14+/CD16-; CD14++/CD16-; CD14+/CD16+; and CD14++/CD16+. The percentage CCR2+ monocytes present in each of the respective monocyte sub-populations were identified using CD195 (CCR2) PE vs SS Log two-parameter plots gated on the respective subpopulations. The overlay plots within the black bordered square indicates the strategy used to determine CCR2 expression of the different monocyte subsets. The negative/positive staining boundaries were determined based on the negative expression of CCR2 by CD16++/CD14- neutrophils (indicated in red in the overlay plots). The CCR2+ populations are indicated in blue. en_ZA
dc.description Additional file 3: Supplementary Figure S3. Maternal intake of estimated average requirements for macronutrients, vitamins and minerals for mothers who report on household food security circumstances. Maternal reports of food insecurity did not associate with intake levels of macronutrients or minerals. Maternal reports of experiencing food runout or inability to afford balanced meals associated with lower intake of vitamin B12 (p=0.01; p=0.04). Many women, irrespective of food security reports, are at risk of inadequate macronutrient, vitamin and mineral intakes. Percent intake of EARs for 36 nutrients were calculated for lactating women 14-18, 19-30 or 31-50 years of age [37]. Calculations for EAR for total protein considered maternal weight at time of dietary recall. Data are % intake of EAR reported in maternal dietary recall for macronutrients, *p<0.05 [ANOVA for normal distribution/ equal variance; Kruskal-Wallis/Wilcoxon test for nonparametric data; or Welch’s test for normal data/unequal variance]). CHO = carbohydrates. en_ZA
dc.description Additional file 4: Supplementary Figure S4. Cooccurrence of maternal HIV and food insecurity may increase risk of stunting at birth. Amongst infants whose mothers report worrying about food runout, risk of stunting at birth is greater for HEU compared to HUU infants (e; RR=4.90 [0.76, 31.5], ARD=0.56 [0.17, 0.94], p=0.0498). The red line represents the proportion of infants who had stunting at birth or 12 weeks PP. Mosaic plots are proportion (%) of HUU or HEU infants who have stunting (<-2 SD length-for-age standardised according to WHO child growth standards [28]) at birth and 12 weeks old. HUU = HIV-unexposed, uninfected infant; HEU = HIV-exposed, uninfected infant. RR = Relative risk. ARD = Absolute risk difference. en_ZA
dc.description Additional file 5: Supplementary Figure S5. Food insecurity may associate with low attainment of GMCD milestones for HUU and HEU infants. Infants whose mothers reported household food insecurity did not attain 1-3 month GMCD milestones (A, C, E) for receptive language, large movement, relating and response behaviour or play activities, or 3-5 month GMCD milestones (B, D, F) for fine movement or relating and response behaviour in the same proportion as the international standardization sample. Maternal reports of food insecurity did not associate with risk of not attaining all 1-3 month or 3-5 month GMCD milestones (A-F, [p>0.05], Fisher’s exact 2-Tail). Data are proportion (%) of infants who attained all age-appropriate GMCD milestones. The horizontal dotted line represents the GMCD standardised international sample proportion (85%) of infants who attained all milestones in that age category, when they were in that age range. The numbers underneath the bars represent the number of infants attaining all milestones for each milestone. GMCD = Guide for monitoring child development; HUU = HIVunexposed, uninfected infant; HEU = HIVexposed, uninfected infant. en_ZA
dc.description Additional file 6: Supplementary Table S1. Flow cytometry reagent list (including lasers and detectors used). Supplementary Table S2. Flow cytometry compensation matrix. Supplementary Table S3. Maternal nutrient intakes from one 24-hour dietary recall for mothers with and without HIV who attended follow up. Supplementary Table S4. Maternal nutrient intake from one 24-hour dietary recall for mothers who report experiencing food insecurity compared to those who do not experience food insecurity. en_ZA
dc.description.abstract BACKGROUND : As mother-to-child transmission of HIV decreases, and the population of infants who are born HIV-exposed, but uninfected (HEU) continues to rise, there is a growing need to understand the development and health outcomes of infants who are HEU to ensure that they have the healthiest start to life. METHODS : In a prospective cohort pilot study at Kalafong Hospital, Pretoria, South Africa, we aimed to determine if we could recruit new mothers living with HIV on antiretrovirals (ART; n = 20) and not on ART (n = 20) and new mothers without HIV (n = 20) through our clinics to study the effects of HEU on growth and immune- and neurodevelopment in infants in early life, and test the hypothesis that infants who were HEU would have poorer health outcomes compared to infants who were HIV-unexposed, uninfected (HUU). We also undertook exploratory analyses to investigate relationships between the early nutritional environment, food insecurity and infant development. Infant growth, neurodevelopment (Guide for Monitoring Child Development [GMCD]) and levels of monocyte subsets (CD14, CD16 and CCR2 expression [flow cytometry]) were measured in infants at birth and 12 weeks (range 8–16 weeks). RESULTS : We recruited 33 women living with HIV on ART and 22 women living without HIV within 4 days of delivery from June to December 2016. Twenty-one women living with HIV and 10 without HIV returned for a follow-up appointment at 12 weeks postpartum. The high mobility of this population presented major challenges to participant retention. Preliminary analyses revealed lower head circumference and elevated CCR2+ (% and median fluorescence intensity) on monocytes at birth among infants who were HEU compared to HUU. Maternal reports of food insecurity were associated with lower maternal nutrient intakes at 12 weeks postpartum and increased risk of stunting at birth for infants who were HEU, but not infants who were HUU. CONCLUSIONS : Our small feasibility pilot study suggests that HEU may adversely affect infant development, and further, infants who are HEU may be even more vulnerable to the programming effects of suboptimal nutrition in utero and postnatally. This pilot and preliminary analyses have been used to inform our research questions and protocol in our ongoing, full-scale study. en_ZA
dc.description.department Immunology en_ZA
dc.description.department Obstetrics and Gynaecology en_ZA
dc.description.department Paediatrics and Child Health en_ZA
dc.description.librarian am2021 en_ZA
dc.description.sponsorship The Collaborative Initiative for Paediatric HIV Education and Research (CIPHER); the Faculty of Science, Carleton University; the Canadian Institutes of Health Research (CIHR); a Canadian Graduate Scholarship-Master’s and a Michael Smith Foreign Study Supplement from CIHR. en_ZA
dc.description.uri https://pilotfeasibilitystudies.biomedcentral.com en_ZA
dc.identifier.citation White, M, Feucht, UD & Duffley, E 2020, 'Does in utero HIV exposure and the early nutritional environment influence infant development and immune outcomes? Findings from a pilot study in Pretoria, South Africa', Pilot and Feasibility Studies, vol. 6, no. 192, pp. 1-20. en_ZA
dc.identifier.issn 2055-5784 (online)
dc.identifier.other 10.1186/s40814-020-00725-8
dc.identifier.uri http://hdl.handle.net/2263/80983
dc.language.iso en en_ZA
dc.publisher BMC en_ZA
dc.rights © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License. en_ZA
dc.subject Pilot study en_ZA
dc.subject Neurodevelopment en_ZA
dc.subject Inflammation en_ZA
dc.subject Growth en_ZA
dc.subject Nutrition en_ZA
dc.subject Food security en_ZA
dc.subject Human immunodeficiency virus (HIV) en_ZA
dc.subject Mother-to-child transmission (MTCT) en_ZA
dc.subject HIV-exposed, but uninfected (HEU) en_ZA
dc.subject Antiretroviral therapy (ART) en_ZA
dc.title Does in utero HIV exposure and the early nutritional environment influence infant development and immune outcomes? Findings from a pilot study in Pretoria, South Africa en_ZA
dc.type Article en_ZA


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