The Occurrence Of Monoclonal Antibodies Shrimp Blood Cells And Virus Infection

Crustacean does not have immunoglobulin. Haemocytes, distributed in the whole body by the blood flow, play important roles in the host immune responses including phagocytosis, clotting reaction, encapsulation etc. In the paper the problems on haemocyte genesis and differentiation, and reaction of haemocyte during the virus (WSSV) invaded the shrimp are thoroughly studied. The research was helpful to explain the questions such as crustacean immune mechanism, the relation between circulating haemocyte and hematopoietic cell, the molecular mechanism of WSSV infection, and find effective methods for curing the disease etc.Haemocytes do not have mitosis, so released continuously from haematopoietic tissue. In this paper, by monoclonal antibodies against haemocytes of the shrimp (Penaeus chinensis), using frozen section and indirect immunofluorescent technique (UFA), location of hematopoietic tissue was identified in the postlarva of Penaeus chinensis. The results showed that hematopoietic tissues were located in the anterior part of stomach, upper part of stomach, base of rostrum, the lepis of antenna, the third maxilliped, base of appendages on the cephalothorax etc. Hematopoietic tissues have two different kinds of organized morphologic characters. In one kind hematopoietic tissue was surrounded the vessel, and in the other kind haemocytes were densely packed in small lobules. At the same time difference between circulating haemocyte and hematopoietic cell was compared.Thorough understanding about the molecular mechanism of WSSV pathogenesis and spreading is still urgently needed in aquatic animal disease study. In this paper, using mouse anti-WSSV monoclonal antibodies and rabbit anti-haemocyte of the shrimp (Penaeus chinensis) polyclonal antibodies as primary antibodies, employing alkaline phosphatase (AP) conjugated sheep anti-mouse Ig serum and horseradish peroxidase (HRP) conjugated sheep anti-rabbit Ig serum as secondary antibodies respectively, double-labeled immunohischemical technique was applied to detect WSSV infected haemocyte in diseased shrimp tissue. Two different signals, whichwere brown and blue, reacted with the substrate (DAB and NBT/BCIP) separately, could be observed in the WSSV infected haemocyte. The result showed that in the lightly infected tissues infected haemocytes predominated in all infected cells. However in the heavily infected tissues the number of normal haemocyte decreased obviously and infected cell with strong positive intensity increased, and cell debris and tissue necrosis were observed. It was deduced that haemocyte played an important role in WSSV infection.