家蝇幼虫血细胞类型及免疫功能的初步研究

目的用不同方法观察家蝇3龄幼虫血细胞的形态,并对血细胞进行分类和免疫功能研究,为昆虫血细胞形态、分类及免疫研究提供实验依据。方法 (1)应用姬氏染色结合相差显微镜及荧光染色方法观察家蝇3龄幼虫血细胞形态,并对家蝇3龄幼虫血细胞进行分类;(2)观察家蝇3龄幼虫感染大肠杆菌后不同时间血细胞总数(THC)、各类血细胞数量(DHC)及形态的变化;(3)应用倒置显微镜观察家蝇3龄幼虫离体血细胞感染大肠杆菌后的形态变化;(4)采用酶细胞化学技术测定感染前后家蝇3龄幼虫血细胞中ACP、POD活性的变化。结果 (1)家蝇3龄幼虫血细胞可分为原血胞、浆血胞、粒血胞、珠血胞、类绛血胞5类,其中浆血胞又分为大核浆血胞和小核浆血胞两种;(2)感染后各时间组血细胞总数、浆血胞和粒血胞数量均显著升高,且浆血胞和粒血胞聚集成堆,出现细胞变形、空泡等形态变化;感染后16h、24h组的珠血胞数显著升高;原血胞和类绛血胞数量和形态无明显变化;(3)家蝇幼虫离体血细胞感染大肠杆菌后粒血胞周围见大量细菌聚集,浆血胞、粒血胞聚集成团将细菌包裹形成包囊,未见原血胞、珠血胞、类绛血胞形态的变化;(4)感染后浆血胞和粒血胞中ACP、POD的活性增强,感染前后原血胞、珠血胞、类绛血胞中均未见ACP、POD的阳性反应物。结论通过3种方法能很好地将家蝇3龄幼虫血细胞分为5类,其中浆血胞和粒血胞是家蝇幼虫参与免疫反应的主要细胞类型,珠血胞不参与感染后的早期细胞免疫反应。

Study on Hemocytes Classification and Immunologic Function of Musca domestica Larva

Objective This study is designed to observe the hemocytes morphous and classification of the 3th stage larva of the Musca domestica by different methods,and to investigate the immunologic function of hemocyte,which provide the experiment proof for the morphous,classification and immunifaction of hemocytes in insects.Methods(1) The hemocyte morphous of the 3th stage larva of Musca domestica was observed and classified by Giemsa staining combined with phase contrast microscope and fluorescent staining.(2) The number changes of total hemocytes(THC),the number of the different hemocytes(DHC) and the hemocytes morphous at different times after being infected by Escherichia coli were examined.(3) The changes of the hemocytes morphous for the Musca domestica larva in vitro infected by E.coli were observed by inverted microscope.(4) The activities of ACP and POD in hemocyte of Musca domestica larva after being infected by E.coli were assayed by enzyme cytochemistry method.Results(1)The hemocytes of the 3th stage larva of Musca domestica could be divided into 5 types:prohemocyte,plasmatocyte,granulocyte,sphrulocyte and oenocytoid,of which the plasmatocyte could be divided into 2 kinds:big nucleus hematocyte and micronuclei hematocyte.(2) The number of total hemocytes(THC),plasmatocytes and granulocytes of the all time groups after infection were significantly increased,and the plasmatocyte and granulocyte after infection aggregated,and the cytomorphosis and vacuole also occurred;the number of sphrulocytes of 16 h and 24 h groups after infection were significantly increased;but the number and morphous of prohemocytes and oenocytoids were not obviously distinguished.(3) E.coli were injected into the hemocytes of the Musca domestica in vitro,a great quantity bacterium to gather granulocyte surrounding,the plasmatocyte and granulocyte aggregation to parcel bacteriums lead to encystment,but the changes of hemocytes morphous are not found in the prohemocyte,sphrulocyte or oenocytoid.(4) The activities of both ACP and POD in the plasmatocyte and granulocyte after infection were much higher than those of the normal groups,the staining of ACP and POD in the prohemocyte,sphrulocyte and oenocytoid were exhibited the negative reaction.Conclusion The hemocytes of the 3th stage larva of Musca domestica are divided into five types by three methods finely,and the plasmatocyte and granulocyte are primary participant of the cell immunity in the Musca domestica larva,but the sphrulocyte are not involved in the early stage cell immune reactions.