The expressed TCRß CDR3 repertoire is dominated by conserved DNA sequences in channel catfish.

We analyzed by high-throughput sequencing T cell receptor beta CDR3 repertoires expressed by αß T cells in outbred channel catfish before and after an immunizing infection with the parasitic protozoan Ichthyophthirius multifiliis. We compared CDR3 repertoires in caudal fin before infection and at three weeks after infection, and in skin, PBL, spleen and head kidney at seven and twenty-one weeks after infection. Public clonotypes with the same CDR3 amino acid sequence were expressed by αß T cells that underwent clonal expansion following development of immunity. These clonally expanded αß T cells were primarily located in spleen and skin, which is a site of infection. Although multiple DNA sequences were expected to code for each public clonotype, each public clonotype was predominately coded by an identical CDR3 DNA sequence in combination with the same J gene in all fish. The processes underlying this shared use of CDR3 DNA sequences are not clear.

B cell memory following infection and challenge of channel catfish with Ichthyophthirius multifiliis.

B cell responses in channel catfish to infection with the parasitic ciliate Ichthyophthirius multifiliis were followed for 3 years. High titers of serum IgM antibodies recognizing I. multifiliis immobilization antigens were present 5weeks after immunizing infection, but by 1 year titers were at low or undetectable levels. Two to three years after infection the numbers of antibody secreting cells recognizing immobilization antigens in skin and head kidney of immune fish had decreased to the level found in uninfected controls. Challenge of immune fish showed they remained immune and that the numbers of antibody secreting cells recognizing immobilization antigens increased in skin but not head kidney. This suggests that antigen-specific memory B cells persisted for 3 years after infection and upon challenge differentiated into antibody secreting cells that localized in skin. Our results suggest that humoral immunity in channel catfish is maintained through IgM(+) memory B cells.

Cross-immunity and antibody responses to different immobilisation serotypes of Ichthyophthirius multifiliis.

Vaccination of channel catfish with either of two serotypes of the parasitic ciliate Ichthyophthirius multifiliis conferred protection against challenge infection by either serotype. Fish were vaccinated by intracoelomic injection with live theronts of isolate G5 (serotype D) or isolate G12 (a new serotype), which express different surface immobilisation antigens. Vaccination with live G12 theronts conferred complete protection against subsequent challenge by both serotypes while vaccination with G5 theronts elicited only partial protection against both serotypes. Vaccination with trophont lysates did not protect against challenge infection. Sera from vaccinated fish were tested in immobilisation assays, ELISAs, and Western blots. Serum antibodies recognised only immobilisation antigens of the serotype used for vaccination in immobilisation assays or on Western blots. No antigens common to both serotypes were identified by Western blots. In contrast, serum antibodies bound antigens in cell lysates from both serotypes by ELISA, demonstrating that antibodies recognising both serotypes are produced in response to infection, which presumably confer observed cross-serotype protection.

Differences in virulence between two serotypes of Ichthyophthirius multifiliis.

Naive channel catfish Ictalurus punctatus were infected by 2 isolates of the parasitic ciliate Ichthyophthirius multifiliis that differed in virulence. The isolates, NY1 and G5, Serotypes A and D, respectively, express different surface immobilization-antigens. The virulence of the 2 isolates was compared using tail-fin infections to quantitate parasite numbers and by analysis of the survival of infected fish. Although NY1 infected fish at a lower level than G5, all NY1-infected fish died, but 51% of G5-infected fish survived. The greater virulence of NY1 is apparently a consequence of its shorter life cycle, which results in overwhelming reinfection of fish before they can develop a protective immune response. This report represents the first experimental evidence for differences in virulence between serotypes of I. multifiliis.