In contrast to mammalian systems, avian species lack a resident or harvestable macrophage population in the abdominal exudate. Peritoneal macrophages in the chicken can be elicited if an inflammatory agent such as sephadex is injected. This study examines the kinetics of different macrophage populations, derived by different methods of isolation and from different hosts, with respect to the elicited oxidative burst upon infection with host-adapted Salmonella serotypes.
The nature of the oxidative burst elicited by murine and avian-derived and cell line macrophages was determined after stimulation with phorbol myristate (PMA), zymosan A, and Salmonella serotypes. Both murine and chicken peritoneal macrophages, chicken blood monocytes and corresponding cell lines, J774A.1 and HD-11, were unable to produce a detectable chemiluminescent (CL) response after interaction with Salmonella using the luminescent probe luminol. However, both PMA and zymosan A induced a CL response in all cell types, with PMA eliciting a higher and earlier peak response (pkH) than zymosan A. Lucigenin-enhanced CL in both murine and chicken macrophages was achieved with PMA, zymosan A and Salmonella serotypes. In this case, zymosan A induced higher responses than PMA. In the peritoneal macrophages of both hosts, there were no significant differences in the oxidative burst induced by the different Salmonella serotypes. However, the J774A.1 (murine) cells demonstrated significant differences, with S. enterica serotype Choleraesuis (S.choleraesuis and S. gallinarum producing the highest response. In the HD-11 (chicken) cells, S. choleraesuis and S. dublin elicited the higher CL. With both cell lines, S. abortusovis failed to induce an appreciable CL response.
In these experiments it was demonstrated that oxidative burst was not detectable in monocytes/macrophage populations using luminol, which suggests a link to the lack of a myeloperoxidase system in these cells. Lucigenin-enhanced CL appeared independent from the myeloperoxidase system, indicating production of another oxidative species compared with luminol. No discernable effect of host specificity with regard to Salmonella serotype and respective host was seen in host-derived or cell line macrophages, and cell line macrophages displayed altered functional characteristics with regard to oxidative burst in comparison with their primary counterparts. 相似文献
Chinese native chicken breeds provide useful resources for the study of genetic diversity. In this study, the alleles of CD8 alpha and CD3d cDNA from Chinese native and introduced western breeds of chicken were analyzed at the sequence level. Six alleles were found, due to 13 amino acid replacements in the extracellular domain of the CD8 alpha sequence. There were four alleles detected in the Chinese strains, and alleles 5 and 6 were identified for the first time. Allele 6 was shared by Langshan, Beijing Fatty and Recessive White Feather chickens. Allele 2, found in the Bigbone strain, was the same as that found in the Leghorn strain H.B15.H7, and allele 3 in the Xianju breed was also the same as in the Leghorn strain H.B15.H12. Two Leghorn lines (RPL line 7 and AY519197) and the Camellia possessed an allele (alleles 1, 4 and 5), respectively, that was not found in the other lines. Nine out of 13 amino acid replacements were situated in the putative major histocompatibility complex (MHC) class I binding CDR1 (positions 30, 33 and 34), CDR2 (positions 58, 62, 63 and 65) and CDR3 (positions 90 and 106). Except for the Xianju breed, the CD8 alpha cDNA of Chinese native chicken breeds shared high homology. Two alleles were found in CD3d. Three additional nucleotides were found at positions 64, 65 and 66 in the newly discovered allele 2. This led to a difference of four amino acids (at residues 22, 23, 24 and 25) in the extracellular domain of CD3d cDNA from the Gushi, Recessive White Feather and ISA chickens compared with these of the White Leghorn and T11.15 (NM_205512). Five hybridoma clones (1C9, 1H5, 4B11, 6G5 and 13C5) against chicken CD8 alpha were generated by DNA immunization. Two monoclonal antibodies (mAbs), 6G5 and 4B11, showed reactivity to the splenocytes from five Chinese native chicken breeds, the Recessive White Feather chicken and the Leghorn (AY519197), while mAbs 1C9, 1H5 and 13C5 showed no reaction with these breeds. 相似文献