Production of house finch (Carpodacus mexicanus) IgA specific anti-sera and its application in immunohistochemistry and in ELISA for detection of Mycoplasma gallisepticum-specific IgA.
The IgA antibody response plays a vital role in mucosal immunity because it functions to neutralize pathogens at the mucosal surface and thus impedes attachment to underlying tissues. Although the importance of IgA in the mucosal immunity of galliform birds has been established, studies examining IgA-based immunity in passerine birds are lacking, perhaps due in part to the absence of reagents that can detect passerine IgA. A 469 base pair region of the house finch (Carpodacus mexicanus) IgA heavy chain was PCR-amplified from spleen cDNA and sequenced. The predicted amino acid sequence was found to share 55% and 46% identity with the IgA heavy chain of mallard (Anas platyrhynchos) and chicken (Gallus gallus), respectively. The heavy chain fragment was produced using a bacterial expression system and purified. Rabbit anti-sera were generated against the recombinant protein. The anti-sera reacted with a single house finch serum protein ( approximately 50-55kDa) in Western blot. The anti-sera were used to identify plasma cells in the Harderian gland and conjunctiva of house finches with conjunctivitis associated with Mycoplasma gallisepticum infection. The anti-sera were also utilized in an ELISA to detect M. gallisepticum-specific IgA antibodies in lachrymal samples of infected finches.
Grodio
Jessica L
JL
College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, United States.
Buckles
Elizabeth L
EL
Schat
Karel A
KA
D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis*; D000907:Antibodies, Bacterial / Q000032:analysis*; D000918:Antibody Specificity; D001483:Base Sequence; D001715:Bird Diseases / Q000276:immunology / Q000382:microbiology; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay; D046369:Finches / Q000235:genetics / Q000276:immunology*; D006243:Harderian Gland / Q000276:immunology / Q000382:microbiology; D018928:Immunity, Mucosal; D007071:Immunoglobulin A, Secretory / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary; D045686:Mycoplasma gallisepticum / Q000276:immunology*; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
D000888:Antibodies, Anti-Idiotypic; D000907:Antibodies, Bacterial; D017931:DNA Primers; D018076:DNA, Complementary; D007071:Immunoglobulin A, Secretory; D007202:Indicators and Reagents