| The mucosal ecosystem of body,which is consists of mucosa and microecosystem within the epithelium,is the first line to defend the invade of outside antigens.pIgR and FcRn as the pivotal effector molecules for their transport corresponding antibodies to the mucosal surface,neutralize and eliminate pathogens to keeping normal microbial flora and physiological environment within the mucosal ecosystem.pIgR and FcRn product by multiple cells in the airway,digestive tract and urogenital tract etc.will conduct mucosal immunity responses in specific sites.The tonsils(including palatine tonsils,pharyngeal tonsil,lingual tonsil,paraepiglottic tonsils,tonsil of the soft palate,tubal tonsils)as the crucial member of the mucosa-associated lymphoid tissues(MALT),appearing different forms among various animals and playing crucial functions in respiratory tract and all mucosal immune system.The Bactrian camel have all of these 6 tonsils referred above.In order to defined the expression and distribution characteristics of pIgR and FcRn in these tonsils of Bactrian camel and the biological functions of them displaying in local mucosal immunity,at the same time,to explore the species differences of the biological functions of pIgR and FcRn among Bactrian camel and other species.In this thesis,6 clinical normal middle-aged Alashan Bactrian camels(6-16 years)were used as the research subjects.The bioinformatics-related charactristics of pIgR and FcRn ? chain were analysised by bioinformatics method;the coding regions(CDS)of pIgR and FcRn ? chain genes were cloned;the relative expression of PIGR and FCGRT within 6 tonsils were tested by RT-q PCR method;meanwhile,the distribution of pIgR and FcRn positive cells in every region of these 6 tonsils were defined by immunohistochemistry method.The results as follows.(1)expression and bioinformatics analysis of PIGR and FCGRT in different tonsilsReal-time quantitative PCR indicated that the expression of PIGR and FCGRT are highest in pharyngeal tonsil among all 6 kinds of tonsils(p<0.05).Bioinformatics analysis to PIGR and FCGRT showed that pIgR and FcRn ? chain are type I transmembrane protein;the distribution region of pIgR and FcRn ? chain's primary structures are similar to the corresponding parts of mammals like human and bovine;also,the tertiary structures of pIgR and FcRn ? chain between Bactrian camle and human are highly similar.Additionally,homology analysis indicated that the CDS regions of PIGR and FCGRT from clone are highly consensus with the correspomding CDS regions were predicted on Gen Bank.(2)Distribution of pIgR-positive cells in Bactrian camel tonsilsDifferent regions within the same tonsil: palatine tonsils: the follicular region higher than the interfollicular region,and the interfollicular region great than the subepithelial compartment and subreticular epithelium region(p<0.05);paraepiglottic tonsils: the follicular region higher than the interfollicular and subreticular epithelium region,meanwhile,interfollicular and subreticular epithelium region higher than subepithelium region(p<0.05);tonsil of the soft palate: there is no significant differences among follicular,subepithelial compartment and subreticular epithelium region,but all of them higher than the interfollicular region and periglandular zone(p<0.05);tubal tonsils: there is no significant differences among follicular region,interfollicular region and periglandular region(p>0.05),but their all higher than subepithelial compartment(p<0.05).Different tonsils within the same regions: follicular region: paraepiglottic tonsils higher than palatine tonsils and tubal tonsils(p<0.05)which have no significant among these 3 kinds of tonsils;interfollicular region: paraepiglottic tonsils and pharyngeal tonsil(no significant difference)higher than palatine tonsils and tubal tonsils(no significant difference),also,palatine tonsils and tubal tonsils higher than tonsil of the soft palate(p<0.05);subepithelial compartment: pharyngeal tonsil and tonsil of the soft palate(no significant difference)higher than palatine tonsils,lingual tonsils,paraepiglottic tonsils and tubal tonsils(no significant difference among these 4 tonsils)(p<0.05);subreticular epithelium region: paraepiglottic tonsils and tonsil of the soft palate(no significant difference)higher than palatine tonsils(p<0.05);periglandular zone: pharyngeal tonsil and tubal tonsils(no significant difference)higher than tonsil of the soft palate(p<0.05).(3)Distribution of FcRn-positive cells in Bactrian camel tonsilsDifferent regions within the same tonsil: palatine tonsils: subreticular epithelium region higher than other 3 regions(p<0.05);pharyngeal tonsil: follicular region,interfollicular region and periglandular region(no significant difference)are all higher than subepithelial compartment(p<0.05);paraepiglottic tonsils: follicular region and subreticular epithelium region higher than interfollicular region and subepithelial compartment parallelly(p<0.05);tonsil of the soft palate: follicular region,interfollicular region and subreticular epithelium region(no significant difference among them)are higher than subepithelial compartment and periglandular region(no significant difference between them)(p<0.05);tubal tonsils: follicular region and periglandular region(no significant difference between them)higher than interfollicular region and subepithelial compartment(no significant difference between these 2 regions)(p<0.05).Different tonsils within the same regions: follicular region: palatine tonsils,paraepiglottic tonsils and tonsil of the soft palate(no significant difference among them)higher than pharyngeal tonsil and tubal tonsils(no significant difference between them)(p<0.05);interfollicular region: pharyngeal tonsil,paraepiglottic tonsils and tonsil of the soft palate(no significant difference among these 3 tonsils)are all higher than palatine tonsils and tubal tonsils(no significant difference between these 2 tonsils)(p<0.05);subepithelial compartment: palatine tonsils,lingual tonsils and paraepiglottic tonsils(no significant difference among these 3 tonsils)are all higher than pharyngeal tonsil,tonsil of the soft palate and tubal tonsils(no significant difference among these 3 tonsils)(p<0.05);subreticular epithelium region: palatine tonsils and paraepiglottic tonsils(no significant difference between these 2 tonsils)are all higher than tonsil of the soft palate(p<0.05);periglandular region: there are no significant differences among pharyngeal tonsil,tonsil of the soft palate and tubal tonsils(p>0.05).(4)Distribution density of pIgR and FcRn-positive cells in the same region of Bactrian camel tonsilspalatine tonsils: FcRn-positive cells are all higher than pIgR-positive cells in follicular,interfollicular,subepithelial compartment and subreticular epithelium region(p<0.05);pharyngeal tonsil: FcRn-positive cells higher than pIgR-positive cells in subepithelial compartment,but there are no significant differences in other regions;paraepiglottic tonsils: there are no significant differences of positive cells density in each region(p>0.05);the tonsil of the soft palate and lingual tonsils: there is no significant differences of positive cells' density in every region(p>0.05).The above results confirmed that the inductive site of different tonsils may vary during mucosal immune responses,note that the inductive mechanism of mucosal immune responses of different tonsils may different;the dominant distribution of pIgR and FcRn positive cells in every region may be anatomical location-oriented;the high homology of the nucleotide and amino acid sequences of pIgR and FcRn ? chain in genus and the subgenus species reflect the similar biological functions of pIgR and FcRn among Bactrian camle,human,bovine,rodent and other mammals. |
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