OBJECTIVE: In sub-Saharan Africa, 10-40% of childbearing age women are seropositive for HIV. Seronegative neonates born to HIV infected women are at risk for HIV vertical transmission and/or other acquired infections associated with feeding mode used—breastfeeding, formula, or mixed. If breast-fed, the risk of infant conversion to HIV positive status by 24 months of age is greater than 36%: if formula-fed, the risk is over 20. Even so, the risk of mortality by 24 months of age is similar for both feeding groups (24% vs. 20%) due to increased non-HIV infection rates of formula-fed infants. When formula is not freely available to resource-poor women, infant mortality increases three-fold as a result of infection and malnutrition leading WHO/CDC to recommend breastfeeding by HIV seropositive mothers. Of those infants who acquire HIV infection, about 50% succumb to the disease in the first 18 months of life. While use of anti-retrovirals for women and children shows promise, maternal and child mortality remains high, with some areas of Africa projected for zero population growth. A low-cost approach to infant feeding that prevents breastmilk HIV transmission and enteric and respiratory infections associated with formula use would increase survival of infants born to HIV infected mothers. Post-childbearing age women in sub-Saharan Africa have low rates of HIV infection and typically function as extended family caregivers, including surrogate nursing. We therefore postulate that this group of women could supplement the feeding of infants born to seropositive mothers with minimal risk of HIV transfer while providing the immune and nutritional components that enhance infant survival. The purpose of this Phase I study was to examine the immunological components in nipple aspirate fluid (NAF) from older, post-weaned African women and evaluate the feasibility of using NAF as a supplemental feed for HIV seronegative infants. DESIGN: A descriptive and relational design was implemented to examine the immunological components of older women’s NAF as part of a larger parent study on breast cancer prevention. POPULATION, SAMPLE, SETTING, YEARS: In August 2000, non-lactating women (N=48) between 35 and 70 years of age or older and not pregnant were selected from two rural villages in coastal Kenya. These women experienced youthful multiparity and extensive lactation histories, including some previous surrogate nursing. Selection criteria included overall “healthy” with no hypertension, cancer, or “wasting disease” symptoms. VARIABLES STUDIED: A health interview, pregnancy test, clinical breast examination, anthropometric assessment, and NAF aspiration were conducted. Peptides and parent proteins biomarkers of immune function in NAF were evaluated with mass spectrometric analysis. METHODS: Women were recruited at two rural health clinic settings in coastal Kenya. Swahili language informed consent and study instruments were completed. Following a clinical breast examination and a 5-minute self-breast massage, heating pads were placed on the breast and held in place with kanga cloth. NAF,which is is produced by the apocrine, mucosa-generating surface of the breast ductal system, was aspirated bilaterally using a patented nipple fluid aspiration system and stored at -70C. Quantity of samples of NAF ranged from <10 µl to >250 µl. In solution, tryptic digestion of the NAF proteins was followed by tandem mass spectrometric analysis of the peptides on a Thermofinnigan LCQ Deca mass spectrometer. Identification of the parent proteins was undertaken using Sequest analysis software. At least 3 separate peptide matches with Sequest Xcorr values above 1.5 (one of which had to be above 2.0) were required to confirm the presence of the parent protein in the NAF sample. FINDINGS: Proteomic analysis of pooled NAF samples indicated that the most abundant proteins were immunoglobins, accounting for ~40% of the total protein content. In addition, immune complements C3, C4 as well as several forms of casein and lactotransferrin were identified. CONCLUSIONS: NAF from post-childbearing African women contained several components of innate immunity typically found in breast milk. Future research will focus on the cost benefit and selective anti-infectivity properties of NAF as a supplemental feed. IMPLICATIONS: This is a first step to address a 21st Century problem with an old solution by re-formulating the use of culturally-accepted surrogate lactation. Surrogate NAF as an immune complement-rich supplement has potential to reduce health risks of African children born to HIV infected mothers. Disparity in child mortality in sub-Saharan Africa warrants novel approaches to ensure a healthy next generation.
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