Production and characterisation of an antiserum raised against recombinant rainbow trout (Oncorhynchus mykiss) MHC class II beta-chain (MhcOnmy-DAB)

Lierop, M.J.C. van; Knight, John; Secombes, Chris J.; Hermsen, Trudi; Groeneveld, A.; Stet, R.J.M.

doi:10.1006/fsim.1997.0129

Cited by 12 publications

(9 citation statements)

References 23 publications

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“…However, after digestion with endoglycosidase H and PNGase F, PaMHCIIα in ayu MO/MФ exhibited the expected, lower MW. In accordance with our study, the MW of native MHC class II α from rainbow trout gills and PBLs was approximately 32 kDa, higher than the aa-derived MW; after deglycosylation, MW was around 29 kDa (van Lierop et al, 1998;Nath, Kales, Fujiki, & Dixon, 2006).These results demonstrated that PaMHCIIα was definitely N-glycosylated and constituted about 10% of the glycoprotein's total mass. Notably, in channel catfish and stickleback, the molecular mass of native MHC class II α matched completely with the MW calculated from the aa sequence, confirming that glycosylation sites are absent (Thankappan, Fuller, Godwin, Kearse, & McConnell, 2006;Scharsack, Kalbe, & Schaschl, 2007).…”

Section: Discussionsupporting

confidence: 90%

“…Antibodies against MHC class II α are available to map MHC classII distribution and glycosylation in cells/tissues of several fish species (Nath, Kales, Fujiki, & Dixon, 2006 (Thankappan, Fuller, Godwin, Kearse, & McConnell, 2006), rainbow trout (Oncorhynchus mykiss) peripheral bloodleucocytes (PBLs) (van Lierop et al, 1998), along with other tissues (e.g., head kidney, gill, spleen, hindgut, liver, and muscle) (Nath, Kales, Fujiki, & Dixon, 2006;Scharsack, Kalbe, & Schaschl, 2007). The protein class's native form is N-glycosylated (van Lierop et al, 1998;Nath, Kales, Fujiki, & Dixon, 2006).…”

Section: Introductionmentioning

confidence: 99%

“…The protein class's native form is N-glycosylated (van Lierop et al, 1998;Nath, Kales, Fujiki, & Dixon, 2006). Ayu (Plecoglossus altivelis) is aneconomically important teleost widely cultured in East Asia (Japan, China, and Korea).…”

Section: Introductionmentioning

confidence: 99%

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Li¹,

Ding²,

Chen³

et al. 2018

Turk. J. Fish. Aquat. Sci.

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The MHC (major histocompatibility complex) class II family plays a major role in vertebrate immune systems and is normally found only on antigen-presenting cells. In this study, we identified the gene encoding the α (PaMHCIIα) chain of MHC class II molecules in ayu (Plecoglossus altivelis). Phylogenetic analysis showed that PaMHCIIα was tightly grouped with MHC class IIα in rainbow smelt (Osmerus mordax) and salmon. PaMHCIIα transcripts were detected in all tested tissues, with high levels in the spleen, head kidney, and gill; expression increased significantly upon Vibrio anguillarum infection. PaMHCIIα mRNA expression was also up-regulated in monocytes/macrophages (MO/MФ) after in vitro stimulation with V. anguillarum. We prepared antibodies for detecting N-glycosylation of PaMHCIIα or neutralizing PaMHCIIα on ayu MO/MФ. Western blot analysis showed that native PaMHCIIα was indeed N-glycosylated. Moreover, PaMHCIIα neutralization not only significantly inhibited ayu MO/MФ phagocytosis, but also reduced the ability of V. anguillarum to induce MO/MФ-cytokine expression. Overall, these findings reveal that the important role of PaMHCIIα against bacterial challenge, and it likely contributes to pathogen responsiveness in ayu MO/MФ.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

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Li¹,

Ding²,

Chen³

et al. 2018

Turk. J. Fish. Aquat. Sci.

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“…Conforming to our observations, van Lierop et al [20] found that a band in muscle tissue reacted with the antiserum. The molecular weight of this band also appears to be somewhat higher than the band size that we estimated in our blots, indicating that the discrepancy between van Lierop et al's [20] and our molecular weight estimates is attributable to some experimental inaccuracy. There is no reason to believe that the antisera recognise di#erent molecules.…”

Section: Identification Of Mhc Class II Chain In Tissue Extractssupporting

confidence: 78%

“…The theoretical molecular weight of the entire mature MHC class II chain polypeptide in Atlantic salmon is approximately 25 kDa. In comparison, van Lierop et al [20] found that a 34 kDa band reacted with their antiserum IDENTIFICATION OF MHC CLASS II POSITIVE CELLS against the recombinant rainbow trout (Oncorhynchus mykiss) MHC class II chain. After deglycosylation, the band size was reported to be reduced to 31 kDa.…”

Section: Identification Of Mhc Class II Chain In Tissue Extractsmentioning

confidence: 96%

Production of rabbit antisera against recombinant MHC class II β chain and identification of immunoreactive cells in Atlantic salmon (Salmo salar)

Koppang

Hordvik

Bjerkås

et al. 2003

Fish & Shellfish Immunology

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Major histocompatibility class II genes in rainbow trout (Oncorhynchus mykiss) exhibit temperature dependent downregulation

et al. 2006

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Major histocompatibility (MH) class II receptors are expressed on the surface of specialized antigen-presenting cells in vertebrate immune systems. Their function is to present peptides derived from exogenous pathogens to CD4+ T cells. Variation in the level of expression of these genes has been linked to pathogenesis in various diseases. Very little has been published on the function of MH class II receptors in teleost fish to date. In this study, we have produced polyclonal antibodies recognizing MH class II alpha and beta proteins of rainbow trout and employed them to characterize the expression pattern of these genes. Deglycosylation using N-glycosidase F and endoglycosidase H showed that MH class II alpha is glycosylated in rainbow trout. MH class II beta was also found to be glycosylated as reported previously. Results from Northern blotting revealed that the expression of these genes was not affected by exposure of rainbow trout to temperature of 5 degrees C. However, at 2 degrees C, downregulation of MH class II alpha and beta genes was evident at both the mRNA and protein levels as assessed by Northern and Western blotting, respectively. Because MH class II antigens play an important role in generating an immune response to bacterial and fungal pathogens, downregulation of these genes at low temperature could account for the susceptibility of fish to low temperature-related diseases such as bacterial cold-water disease and winter saprolegniosis.

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Production and characterisation of an antiserum raised against recombinant rainbow trout (Oncorhynchus mykiss) MHC class II beta-chain (MhcOnmy-DAB)

Cited by 12 publications

References 23 publications

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Production of rabbit antisera against recombinant MHC class II β chain and identification of immunoreactive cells in Atlantic salmon (Salmo salar)

Major histocompatibility class II genes in rainbow trout (Oncorhynchus mykiss) exhibit temperature dependent downregulation

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