The development of an ELISPOT assay to quantify total and specific antibody-secreting cells in dab Limanda limanda (L.)

Interaction between the blood fluke,Sanguinicola inermisand humoral components of the immune response of carp,Cyprinus carpio

“…Within the physiological range of a particular teleost species it is widely accepted that higher environmental temperatures enhance the magnitude and timing of antibody production (e.g. Rijkers, Frederix-Wolters & van Muiswinkel, 1980 ;Secombes et al 1991). It is of interest that previous studies by Richards et al (1996 a) have also revealed that carp lymphocytes cultured with S. inermis extracts were also temperature sensitive i.e.…”

Quantitation of antibody secreting cells in high and low antibody responder inbred carp (Cyprinus carpioL.) strains

“…Today, a more sensitive assay for the enumeration of individual ASC, the ELISPOT assay, has been developed, based upon immuno-enzyme technology (Czerkinsky et al, 1983;Sedgwick & Holt, 1983). This technique has already been used for detection of ASC in fish such as channel catfish (Ictalurus punctatus: Waterstrat et al, 1991), dab (Limanda limanda: Secombes et al, 1991), rainbow trout (Onchorhynchus mykiss: Davidson et al, , 1993Siwicki & Dunier, 1993), roach (Rutilus rutilus: Aaltonen et al, 1994) and carp (Cyprinus carpio: Koumans-van Diepen et al, 1995), immunised with di#erent antigens. Here, we examined possible quantitative di#erences in ASC, both after primary and secondary immunisation with DNP-KLH, that may explain the selected di#erences in serum antibody production.…”

Modulation of dab ( , L.) macrophage respiratory burst activity

“…These studies have shown that dab possess a number of nonspecific humoral defences, such as serum anti-protease and lysozyme activity (Secombes et al, 1991bTahir et al, 1993). In addition, kidney and spleen leucocyte suspensions have been used to demonstrate nonspecific cellular responses such as phagocytic activity, respiratory burst activity, bactericidal activity and leucocyte locomotion (Secombes et al, 1991aTahir et al, 1993;Pulsford et al, 1994).…”

“…In addition, kidney and spleen leucocyte suspensions have been used to demonstrate nonspecific cellular responses such as phagocytic activity, respiratory burst activity, bactericidal activity and leucocyte locomotion (Secombes et al, 1991aTahir et al, 1993;Pulsford et al, 1994). Of the specific immune responses examined to date, it is clear that dab are capable of secreting specific antibody, that immunoglobulins represent a major serum protein (present at 10-15 mg ml 1 ) and that the kidney is the major site of antibody secreting cells (Secombes et al, 1991a(Secombes et al, , 1991b. Despite these studies, virtually nothing is known about the responses of individual leucocyte populations, since the above assays were carried out using unfractionated leucocyte suspensions, although it could be argued that B-cell responses were being studied with respect to the antibody data.…”