Chain Sera Research Articles

Preparation of anti-bovine and porcine μ chain sera

Absorption of anti-L chain antibodies contained in anti-IgM sera immunized with purified bovine and porcine IgM was undertaken by affinity chromatography on a column of Sepharose 4B, coupled with the normal follicular fluids of the respective species as an immunosorbent. Anti-L chain antibodies in both anti-IgM sera were completely absorbed without incurring reduction of the antibody titer, and anti-bovine and anti-porcine μ chain sera were prepared.

Attachment of serum non-antibody glycoproteins to the Fusobacterium nucleatum found in periodontal disease

Fourteen of 15 human sera displayed inhibition of the characteristic haemagglutination (HA) of oral strains of Fusobacterium nucleatum in a microtitre haemagglutination inhibition (HI) test. Chromatography of a gamma globulin pool revealed the activity in the fractions eluting with IgM and IgA; absorption procedures with anti-human μ and α chain sera did not remove the activity. Complete saturation of the serum with (NH 4) 2SO 4 produced a floating mucinous fraction displaying HI activity. Sepharose 4B Chromatography of glycoprotein preparations gave mol. wt in excess of 400,000; antibody was not present. Glycoprotein from sera of various blood types aggregated whole F. nucleatum cells. EDTA and galactose inhibited this aggregation whereas glucose did not inhibit. Thus serum glycoproteins containing galactose probably bind to the surface of the F. nucleatum and this binding may influence colonization in the gingival sulcus.

Dissociation of lymphocyte activating determinants from recognition sites in mouse MLR.

PREVIOUSLY, we were able to block mixed lymphocyte reactions (MLR) in man using anti-light chain antibody1 but we have been unable to confirm this observation in subsequent studies using other anti-light chain sera. We have now found that MLR in the mouse is unaffected by a variety of anti-immunoglobulin sera but using alloantisera it has become evident that the stimulator cells, but not the responder lymphocytes, can be blocked.

Effects of anti-immunoglobulin sera on migration inhibition mediated by human lymphocytes.

AbstractMigration inhibition experiments were performed with mixtures of sensitized human lymphocytes and guinea pig peritoneal exudate cells. Treatment of human lymphocytes with a rabbit anti‐human light chain serum (ALCS) abolished their capacity to cause antigen‐induced migration inhibition, whereas treatment with anti‐heavy chain sera did not. The specificity of ALCS was confirmed by absorption studies. The finding that anti‐heavy chain sera did not influence migration inhibition is consistent with other evidence that B cells are not directly involved in the migration inhibition process. The results provide indirect evidence that the — presumably thymus‐dependent — lymphocytes primarily involved in the migration inhibition process carry light chain determinants on their membrane.

On the mechanism of the ring zone effect obtained with the mixed haemadsorption technique. Studies with defined model systems.

Studies on the ring zone effect obtained by the mixed haemadsorption technique on thyroid cultures with sera from patients with thyroid autoimmunity were continued in a model system that permitted a controlled variation of some relatively well defined and simple antigens (human serum albumin, HSA, dinitrophenyl‐human serum albumin, DNP‐HSA). Varying doses of these substances were conjugated to human monolayer cultures by the aid of bis‐diazotized benzidine. Reactions to the antigens were examined by the mixed haemadsorption technique using rabbit antisera against HSA and DNP. Ring zone effects were obtained with anti‐HSA sera only after prolonged immunization producing high anti‐HSA titres and only with a narrow range of relatively low HSA concentrations on the cultures. More antibody was eluted from cultures pre‐incubated with ring zone serum than with corresponding concentrations of filled zone serum. Rabbit ring zone anti‐HSA attached to HSA conjugated cultures was traced more effectively by anti‐gamma and anti‐Fc sera than by anti‐L chain sera, while filled zone anti‐HSA was traced equally effectively by all three types of antiglobulin. Anti‐DNP sera did not produce ring zones with any concentration of DNP‐HSA on the cultures or with any obtainable DNP saturation on the HSA molecules. On the other hand, half saturation or more of DNP on the HSA molecules turned the ring zones obtained with anti‐HSA into filled zones. These results were in agreement with those obtained on thyroid cultures and lend further support to the hypothesis that the ring zones are due to steric hindrance effects on the antibodies in densely crowded clusters of determinants representing several antigen specificities.

Surface immunoglobulin of myeloma cells.

AbstractMouse myeloma cells have been stained in suspension at 0 °C or at room temperature with fluorescein‐labeled rabbit anti‐mouse immunoglobulin sera (F‐RAMIG) or with unlabeled RAMIG followed by fluorescein‐labeled goat anti‐rabbit immunoglobulin sera (F‐GARIG). Tissue culture adapted murine myeloma cells 5563 (IgG2aκ) have been used most extensively in these studies. 95–100 % of the cells show surface fluorescence at both temperatures after staining with F‐RAMIG or with RAMIG followed by F‐GARIG. At 0 °C the staining is diffuse over all the surface, at room temperature the staining is much brighter and patchy. The nature of the H and L‐chains at the surface of the myeloma cell is shown to parallel the type of secretory product. Thus, 5563 cells stain brilliantly with anti‐IgG and anti‐κ chain sera, but not at all with anti‐μ or anti‐α chain sera, which respectively stain TEPC 183 (IgMκ) and MOPC 47A (IgAκ) cells brilliantly. Also, MOPC 315 cells (IgAλ) do not stain with antisera against 5563 myeloma protein, but do stain with antisera against MOPC 47A and MOPC 104E myeloma proteins.Immediately after staining at 0 °C or room temperature, the fluorescence is located over the whole surface. However, if the stained cells are incubated at 37 °C in tissue culture medium, the stain gradually localizes to one pole of the cell such that by 5 h, about 50 % of the cells show staining localized to one hemisphere. The presence of the RAMIG‐antigen complex seemed to have little or no effect on the rate of secretion of myeloma protein during a 5 h period. The results are discussed in relation of the findings of others and to the significance of the surface immunoglobulin of myeloma cells.

Subclasses of porcine immunoglobulins G. Production of subclass specific antisera with S-sulfonated gamma chains.

AbstractUsing antisera made against porcine S‐sulfonated heavy chains two subclasses of porcine IgG could be distinguished on the basis of antigenic determinants located on the Fc fragment. The antisera prepared against the porcine gamma chains reacted with other mammalian IgG proteins, but not with avian, amphibian or fish immunoglobulins. Antigenic determinants responsible for cross‐reactivity are further localized on the Fc′ fragment of the porcine IgG molecule. Comparable antisera, specific for porcine light chains, did not cross‐react with IgG from any of the other species tested. Porcine IgG and F(ab′)2 fragment reacted with both the anti‐heavy and anti‐light chain sera, while the Fab fragment reacted only with the anti‐light chain sera. Important antigenic determinants are thus located on the light chains and the Fc portion of the heavy chain of the porcine IgG molecule.