Host specificity and horizontal gene transfer in the MIB-MIP immunoglobulin evasion system in Mycoplasma...

Mycoplasma gallisepticum is a major poultry pathogen responsible for chronic respiratory disease and substantial global economic losses. Its ability to establish chronic infections reflects its effective immune evasion strategies, but the mechanisms underlying this remain poorly understood. Some pathogenic mammalian mycoplasmas use the Mycoplasma Immunoglobulin Binding-Protease (MIB-MIP) system to capture and cleave host immunoglobulins (Ig), but the functionality and host-specificity of this system in M. gallisepticum have not been examined. We aimed to functionally characterise all the MIB-MIP homologues in M. gallisepticum and examine their host specificity. Five putative MIB and five putative MIP genes of M. gallisepticum were cloned, expressed as recombinant GST-fusion proteins, and purified for functional analysis. Immunoglobulin-binding assays showed that all MIB proteins bound both avian and mammalian immunoglobulins, forming stable MIB-Ig complexes, with distinct binding capacities. In contrast, proteolytic assays revealed that only three of the five MIP proteins could cleave avian immunoglobulins, when complexed with any of the five MIBs, generating characteristic Ig heavy-chain fragments. Only one MIP protease showed detectable interaction with mammalian immunoglobulins, indicating strong host specificity of these MIPs and functional specialisation for avian immunoglobulin-cleavage. These results revealed that MIB and MIP proteins of M. gallisepticum are adapted to cleavage of avian immunoglobins, thereby interfering with antibody-mediated host immune responses. Bioinformatic analysis suggested that MIB-MIP homologues are widespread among avian mycoplasmas that share similar hosts, tissue tropisms and transmission patterns, and detected evidence of horizontal gene transfer and recombination, indicating that there have been MIB-MIP evolutionary adaptations among the avian mycoplasmas.