Cross M L, Cupp M S, Cupp E W, Ramberg F B, Enriquez F J
Department of Veterinary Sciences, University of Arizona, Tucson 85721.
J Med Entomol. 1993 Jul;30(4):725-34. doi: 10.1093/jmedent/30.4.725.
The humoral antibody responses to salivary antigens of Simulium vittatum Zetterstedt were investigated in a BALB/c mouse laboratory model. Production of antisera was stimulated by intraperitoneal immunization with salivary gland extract or by feeding flies directly on depilated mice. Antibody responses in these two groups of mice were compared by western blotting, thus characterizing "true" salivary immunogens present in salivary gland extract. Immunized mice developed IgG, IgM, and IgE antibodies which recognized several salivary gland components, ranging in molecular weight between 26 and 67 kDa. Sera from bitten mice recognized fewer antigens, indicating that some components of the salivary gland extract were poorly immunogenic or absent from the saliva secreted during blood feeding. Antisera raised against S. vittatum also were used to identify cross-reactive immunogens and allergens in salivary gland extracts from other New World simuliids (Simulium argus Williston, S. metallicum Bellardi, and S. ochraceum Walker). SDS-PAGE protein profiles indicated a high degree of similarity between salivary gland extract of S. vittatum and S. argus, and several cross-reacting antigens were identified by western blotting. In contrast, protein profiles of S. ochraceum and S. metallicum differed from the former species, both qualitatively and quantitatively. Antisera demonstrated a low degree of cross-reactivity against salivary gland extract of S. ochraceum, whereas no cross-reactivity was detected against S. metallicum. These observations were confirmed using a monoclonal antibody raised against S. vittatum salivary gland extract (designated SVSG.1.F10), which showed cross-reactivity against S. argus but failed to recognize salivary gland components of either S. ochraceum or S. metallicum.
在BALB/c小鼠实验模型中，研究了对条蚋唾液抗原的体液抗体反应。通过用唾液腺提取物进行腹腔免疫或直接让苍蝇叮咬脱毛小鼠来刺激抗血清的产生。通过蛋白质印迹法比较这两组小鼠的抗体反应，从而鉴定唾液腺提取物中存在的“真正”唾液免疫原。免疫小鼠产生了IgG、IgM和IgE抗体，这些抗体识别几种唾液腺成分，分子量在26至67 kDa之间。被叮咬小鼠的血清识别的抗原较少，这表明唾液腺提取物的某些成分免疫原性较差或在吸血过程中分泌的唾液中不存在。针对条蚋产生的抗血清也用于鉴定来自其他新大陆蚋（阿氏蚋、金属蚋和赭色蚋）唾液腺提取物中的交叉反应性免疫原和变应原。SDS-PAGE蛋白质谱表明条蚋和阿氏蚋的唾液腺提取物之间具有高度相似性，并且通过蛋白质印迹法鉴定了几种交叉反应抗原。相比之下，赭色蚋和金属蚋的蛋白质谱在质量和数量上都与前一种不同。抗血清对赭色蚋唾液腺提取物的交叉反应性较低，而对金属蚋未检测到交叉反应性。使用针对条蚋唾液腺提取物产生的单克隆抗体（命名为SVSG.1.F10）证实了这些观察结果，该单克隆抗体对阿氏蚋具有交叉反应性，但未能识别赭色蚋或金属蚋的唾液腺成分。
