Heterogeneity of the light chains of rabbit immunologlobulin G fractions and of a series of antibodies directed towards antigens of deffering complexity

Mozes, E; Robbins, John B.; Sela, Michael

doi:10.1016/0019-2791(67)90185-1

Cited by 22 publications

(5 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The light chains of Ab-VII-peak-b migrated somewhat faster than the light chains of Ab-VII-peak-a, a finding consistent with the mobilities of the intact proteins. Mozes et al (25) have obtained evidence of a similar sort on the mobility of light chains isolated from differently charged "yG-globulin fractions, which were recovered by DEAE-Sephadex. The light chains of A-variant Ab-VII as well as the light chains of other "),-globulins of this serum showed greater heterogeneity.…”

Section: Streptococcal Group A-variant Antisera--depicted Inmentioning

confidence: 84%

The Individual Antigenic Specificity of Antibodies to Streptococcal Carbohydrates

Braun

Krause

1968

The Journal of Experimental Medicine

101

View full text Add to dashboard Cite

Although a single electrophoretically uniform antibody component with specificity for the group carbohydrate may comprise the bulk of the γ-globulin in rabbits immunized with streptococcal vaccines, this is not always the case. Not infrequently, electrophoresis may reveal multiple antibody components. Nevertheless, it has been feasible by various preparative procedures to isolate from a single antiserum at least two antibody components with similar reactivity for the carbohydrate both of which are electrophoretically monodisperse. Light chains from such antibodies reveal a restricted pattern when examined by disc electrophoresis. Antibodies to streptococcal carbohydrates have been examined for their individual antigenic specificity. Goats were immunized with isolated Group C and Group A-variant antibodies raised in rabbits. Individual antigenic specificity of these antibodies was brought out by absorption of the goat anti-antiserum with Fr II of pooled normal rabbit sera. Additional absorption of the goat anti-antisera with Fr II diminished but did not eliminate the reactivity for the homologous antibody. Immunoelectrophoretic studies with papain fragments of purified streptococcal antibodies localized the specificity to the Fab fragment. Specificity was not confined to the isolated light chains of the antibody.

show abstract

Section: Streptococcal Group A-variant Antisera--depicted Inmentioning

confidence: 84%

The Individual Antigenic Specificity of Antibodies to Streptococcal Carbohydrates

Braun

Krause

1968

The Journal of Experimental Medicine

101

View full text Add to dashboard Cite

show abstract

“…The filtered cells as well as those not passed over glass beads were diluted to 2 X 106, 4 X 106, and 8 X 106/ml, and inoculated into syngeneic recipients exposed to 750 R of x-radiation". 24 hr later, each group of recipients was inoculated intraperitoneally with 10 ug of one of the above immunogens in complete Freund's adjuvant (Difco Laboratories). 14 days later (at the time of peak response), the mice were bled and their sera were titered individually for anti-Dnp antibodies by passive microhemagglutination14 with tanned sheep erythrocytes coated with Dnp-rabbit serum albumin and for anti-lysozyme antibodies with lysozyme-coated erythrocytes.…”

mentioning

confidence: 99%

Cellular Aspects of the Inverse Relationship between the Net Charge of Immunogens and of Antibodies Elicited

Sela

Mozes

Shearer

et al. 1970

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Abstract. The results reported establish a cellular basis for the inverse relationship between the net electrical charge of immunogens and of the antibodies provoked by them. Glass bead columns were used to reduce the number of immunocompetent spleen cells preferentially reactive with more acidic immunogens. After a single immunization, titers of antibodies to an acidic dinitrophenylated copolymer of tyrosine, glutamic acid, and lysine (DNP-901) elicited in recipient mice by filtered spleen cells were significantly lower than those generated by unfiltered cells. After secondary stimulation, the major portion of the antibodies provoked by the acidic antigen was found in the more acidic fraction eluted from DEAE-Sephadex, in contrast to the more basic antibodies, of the same specificity, generated by unfiltered spleen cells. Results obtained by transplanting a limiting number of spleen cells indicate a depletion in the number of precursor cells reactive with dinitrophenyl on DNP-901 after glass bead chromatography, whereas there was no change in the response to dinitrophenyl on a basic copolymer, DNP-912, containing the same amino acids in different molar ratios.The inverse relationship between the net electric charge of immunogenic macromolecules and the net charge of antibodies provoked by them has been demonstrated for antibodies produced in rabbits1'2, mice3, goats4, and humans5, and is valid for antibodies of IgG6, IgM7, and, probably, IgEl class. Thus, e.g., antibodies to the acidic diphtheria toxoid and to negatively-charged synthetic copolymers of amino acids appear mainly under the first peak upon fractionation on DEAE-Sephadex 8, whereas antibodies to the basic lysozyme and positivelycharged amino acid copolymers appear predominantly, or exclusively, under the second peak 3.Haptens such as the dinitrophenyl group1 2 or a peptide of L-alanine (unpublished data of Licht, A., B. Schechter, I. Schechter, and M. Sela) will lead to antibodies fractionating under the first or the second DEAE-Sephadex peak, depending on whether they are attached to a negatively or positively charged carrier. The anti-hapten antibodies produced upon immunization with such conjugates differ in their net electric charge, but not in their specificity or affinity, as measured by means of small molecules related chemically to the original haptens. The inverse correlation described depends on the net electric charge of the intact immunogen, and not on the net charge within a limited area around the anti-1288

show abstract

“…A bone marrow-derived population of immunocytes is responsible for the synthesis of specific antibody (18). The antibodies elicited by DNP-901 (T,G,L) and DNP-912 (T, G, L) differ in their net electrical charge, which is reflected in the light chains (44). Therefore, one might have expected that marrow cell fractionation would have affected the anti-DNP responses similarly to that observed by thymocyte fractionation.…”

Section: Discussionmentioning

confidence: 99%

The Role of Thymocytes and Bone Marrow Cells in Defining the Response to the Dinitrophenyl Hapten Attached to Positively and Negatively Charged Synthetic Polypeptide Carriers

Karniely

Mozes

Shearer

et al. 1973

The Journal of Experimental Medicine

View full text Add to dashboard Cite

An inverse relationship exists between the net electrical charge of immunogens and the antibodies they elicit (1). Results of an earlier study have demonstrated that the net charge phenomenon has a cellular basis, since the immune response potential of murine spleen cells to 2,4-dinitrophenyl (DNP) on a negatively charged synthetic polypeptide carrier was reduced by cell fractionation over negatively charged glass beads, whereas the response to the same hapten on a positively charged carrier was unaffected (14). To verify that the net charge correlation is expressed at the cellular level, spleen cells were fractionated over positively charged poly-L-lysine-coated glass bead columns, and their immunocompetence to DNP on positively and negatively charged carriers was tested by cell transfers in irradiated recipient mice. In this case, the fractionated cells showed reduced response potential to DNP on the positively charged carrier only. Thus, the cellular basis of the net charge phenomenon has been demonstrated for both positively and negatively charged immunogens (for the same specificity) by cell separation techniques over columns of opposite charge. In order to establish whether the cell population relevant for the charge properties of immunogens was of thymus or marrow origin, thymocytes and bone marrow cells were selectively passed over positively or negatively charged columns and mixed with unfractionated cells of the complementary type. Transfers of the filtered and unfiltered cell mixtures in irradiated recipient mice immunized with DNP on either a positive or a negative synthetic polypeptide carrier indicated that fractionation of thymocytes, but not of marrow cells, correlated with the spleen population. Thus, thymocytes fractionated over negatively charged columns and mixed with unfractionated marrow cells exhibited reduced response to DNP on the negative carrier, but normal responses to DNP on the positive carrier. The opposite result was obtained when thymocytes were passed over positively charged columns. No effect on the anti-DNP response was detected by filtration of bone marrow cells over columns of either charge. These findings indicate that it is possible to distinguish between thymocytes on the basis of their capacity to react with more acidic or more basic surfaces and that a population of thymus-derived cells may recognize immunogens on the basis of their overall electrical charge. No evidence was found by these techniques that marrow-derived cells contribute to the net charge phenomenon.

show abstract

Heterogeneity of the light chains of rabbit immunologlobulin G fractions and of a series of antibodies directed towards antigens of deffering complexity

Cited by 22 publications

References 32 publications

The Individual Antigenic Specificity of Antibodies to Streptococcal Carbohydrates

The Individual Antigenic Specificity of Antibodies to Streptococcal Carbohydrates

Cellular Aspects of the Inverse Relationship between the Net Charge of Immunogens and of Antibodies Elicited

The Role of Thymocytes and Bone Marrow Cells in Defining the Response to the Dinitrophenyl Hapten Attached to Positively and Negatively Charged Synthetic Polypeptide Carriers

Contact Info

Product

Resources

About