Steven W M, Barron R A, Stewart G L, Seelig L L
Department of Cellular Biology and Anatomy, Louisiana State University Medical Center, Shreveport 71130.
Alcohol Alcohol. 1991;26(5-6):615-25. doi: 10.1093/oxfordjournals.alcalc.a045165.
Significant immunological protection is provided to the newborn by the transfer of maternal leukocytes during nursing. The objective of this study was to determine if ethanol ingestion altered the distribution of T, B and accessory cells in the mammary glands of normal rats or in rats infected with Trichinella spiralis (T. spiralis). In addition, female rats were fed either Lieber/DeCarli regular (18% protein) or higher protein (25% protein) ethanol-containing liquid diets for pregnant or lactating rats or were pair-fed the corresponding isocaloric control diets to assess changes in cellular distribution due to dietary content. In the first experiment (short-term ethanol), animals were placed on either 18 or 25% protein diet (ethanol or pair-fed) on day 1 of pregnancy. In the second experiment (long-term ethanol), animals were placed on 18% protein diet (ethanol or pair-fed) for 30 days prior to mating, mated and then placed on either 25% protein diet (18%/25% animals) or kept on 18% protein diet (18%/18% animals) through pregnancy and lactation. In the third experiment (long-term ethanol, immunized), animals were fed 18%/25% protein diet (ethanol or pair-fed) as in experiment 2 and were infected intragastrically with 1000 T. spiralis larva on day 15 of pregnancy. All animals were sacrificed on day 2 of lactation, the mammary gland removed and weighed. Frozen sections of mammary gland from each animal were incubated with monoclonal antibodies directed against antigens associated with rat T cells (W3/13), suppressor/cytotoxic (OX8) or helper (W3/25) T cell subsets, macrophages (ED2) or rat IgA (B cells) and processed for peroxidase anti-peroxidase (PAP) immunocytochemistry. The total number of antigen positive leukocytes/high powered microscope field (hpf), intraepithelial leukocytes/hpf and alveolar connective tissue leukocytes/hpf was determined for each stain by counting 100 hpf and the data were compared by statistical analysis. Significant differences in mammary gland weight between ethanol-treated and pair-fed animals were observed in the long-term 18%/18% protein diet experiment in which animals fed ethanol had significantly lower mammary gland weights. Ethanol-treated short-term animals maintained on the 18% protein diet through pregnancy, showed decreased numbers of IgA+ B cells and W3/13+ T cells/hpf in the alveolar connective tissue compartment and this decrease was reflected in total cells/hpf. Long-term 18%/18% protein diet, ethanol-fed animal showed increases in total IgA+ B cells and W3/13+ T cells/hpf as compared with pair-fed controls.(ABSTRACT TRUNCATED AT 400 WORDS)
哺乳期间母体白细胞的转移为新生儿提供了重要的免疫保护。本研究的目的是确定乙醇摄入是否会改变正常大鼠或感染旋毛虫的大鼠乳腺中T细胞、B细胞和辅助细胞的分布。此外，给怀孕或哺乳的雌性大鼠喂食Lieber/DeCarli常规（18%蛋白质）或高蛋白（25%蛋白质）含乙醇液体饮食，或配对喂食相应的等热量对照饮食，以评估饮食成分导致的细胞分布变化。在第一个实验（短期乙醇实验）中，动物在怀孕第1天开始食用18%或25%蛋白质饮食（乙醇组或配对喂食组）。在第二个实验（长期乙醇实验）中，动物在交配前30天食用18%蛋白质饮食（乙醇组或配对喂食组），交配后，在整个怀孕和哺乳期间，将其置于25%蛋白质饮食（18%/25%动物组）或继续置于18%蛋白质饮食（18%/18%动物组）中。在第三个实验（长期乙醇实验，免疫组）中，动物如实验2那样喂食18%/25%蛋白质饮食（乙醇组或配对喂食组），并在怀孕第15天经胃内感染1000条旋毛虫幼虫。所有动物在哺乳第2天处死，取出乳腺并称重。将每只动物乳腺的冰冻切片与针对与大鼠T细胞（W3/13）、抑制/细胞毒性（OX8）或辅助（W3/25）T细胞亚群、巨噬细胞（ED2）或大鼠IgA（B细胞）相关抗原的单克隆抗体孵育，并进行过氧化物酶抗过氧化物酶（PAP）免疫细胞化学处理。通过计数100个高倍显微镜视野（hpf）来确定每种染色的抗原阳性白细胞总数/hpf、上皮内白细胞/hpf和肺泡结缔组织白细胞/hpf，并通过统计分析比较数据。在长期18%/18%蛋白质饮食实验中，观察到乙醇处理组和配对喂食组动物的乳腺重量存在显著差异，喂食乙醇的动物乳腺重量明显较低。在怀孕期一直食用18%蛋白质饮食的短期乙醇处理动物中，肺泡结缔组织隔室中IgA+B细胞和W3/13+T细胞/hpf数量减少，这种减少反映在总细胞/hpf中。与配对喂食的对照组相比，长期食用18%/18%蛋白质饮食的乙醇喂养动物中，总IgA+B细胞和W3/13+T细胞/hpf增加。（摘要截短至400字）
