The immune response against anti‐idiotope antibodies II. The induction of antibodies bearing the target idiotope (Ab3β) depends on the frequency of the corresponding B cells

Abstract

In the present study we use two monoclonal anti-idiotope antibodies (Ac38 and Ac146; Ab1) against the germ line-encoded, lambda 1 chain-bearing and (4-hydroxy-3-nitrophenyl)acetyl (NP)-binding antibody B1-8 (Ab1) for the induction of complementary antibodies (Ab3) and ask the question to what extent antibodies bearing B1-8-like idiotopes (Ab3 beta) are represented in the Ab3 population. In this experimental system, Ab3 beta is distinguished from the remaining Ab3 population (Ab3 alpha) by three properties which Ab3 beta may share with B1-8: (a) the binding of NP, (b) the binding to Ac146 (if induced by Ac38) or the binding to Ac38 (if induced by 146) and (c) that they carry lambda 1 chains. Antibodies with all three properties are induced in low amounts by both anti-idiotopes. Also, Ac146 induces only Ab3 alpha (bearing kappa chains and not binding NP and Ac38). In contrast, Ac38 triggers almost exclusively a lambda 1 chain-bearing response, i.e. Ab3 beta. The response has an unusually large size, reaches its maximum after a week and is long-lasting. An analysis at the level of lipopolysaccharide-reactive precursor B cells demonstrates that, in this case, cells expressing Ab3 beta occur at exceedingly high frequency (approximately equal to 10(-3] and are at least 10 times more frequent than cells expressing Ab3 alpha. The high frequency of Ab3 beta-expressing cells correlates with the contribution of several VH, D and J genes to the expression of this particular idiotope. In the case of the Ac146 anti-idiotope antibody, the response is dominated by Ab3 alpha. Ab3 beta represents less than 10% of the total response, reaches maximal levels 2 weeks after immunization and declines rapidly. This correlates with a low frequency of precursor B cells expressing Ab3 beta (approximately equal to 10(-5] and a restriction of the corresponding idiotope to rare VH-D combinations, caused presumably by a stringent contribution of the H chain to this idiotope which covers the NP-binding site. Our data suggest that anti-idiotypic antibodies (Ab2) against Ab1 will preferentially induce antibodies idiotypically related to Ab1 if the corresponding idiotopes are expressed in high frequency in the B cell compartment. This is expected in cases where Ab2 recognizes an idiotope that can be formed by many germ line-encoded VH-D-VL-combinations.

Disparate adjuvant properties among three formulations of “alum”

“…As a comparator, NP 11 -CGG was adsorbed in the same way to Alhydrogel®, an alum standard often used to evaluate new adjuvants[24]. Finally, NP 11 -CGG was also precipitated with alum as a third comparator [19]. C57BL/6 mice were immunized i.p.…”

“…For Imject® and Alhydrogel® adjuvants, antigen/adjuvant preparations followed the methods recommended by the manufacturer; precipitated alum/antigen preparations were generated as described [19]. Briefly, antigen was adsorbed to Imject® alum (Thermo Scientific; 40 mg/ml aluminum hydroxide, 40 mg/ml magnesium hydroxide) in a solution (1 mL volume) of sterile PBS containing 200 μg/ml antigen.…”

“…Antigen/aluminum precipitates were generated as follows [19]: 5 g of Al 2 (SO 4 ) 3 ·(14-18) H 2 0 (EM Science) was dissolved in 50 ml water to make a 10% Al 2 (SO 4 ) 3 solution; 1 ml 200 μg/ml antigen solution was mixed with 1 ml 10% Al 2 (SO 4 ) 3 solution. Subsequently, KOH (1 M) was added drop-wise with constant stirring until pH 6.5.…”

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. I. The architecture and dynamics of responding cell populations.

“…Groups of C57BL/6 and F, mice were immunized with a single intraperitoneal injection of 50 Fig NP-CG/alum . Another group of C57BL/6 mice were immunized with in intraperitoneal injection of 50 hg of the andidiotope mAb, Ac38, coupled to keyhole limpet hemocyanin (Ac38-KLH), and precipitated in alum as described by Tesch et al (35) . For controls, a number of C57BL/6 mice were immunized with 50 hg CG precipitated in alum (CG/alum).…”

Immune response against poly(Glu60, Ala30, Tyr10) (GAT): immunization with monoclonal anti‐idiotypic antibodies leads to the predominant stimulation of idiotypically similar immunoglobulins with anti‐GAT activity

“…This response is expected to consist of immunoglobulin (Ig) idiotypically related to Abl with or without antibody activity (Abl') and of anti-anti-Id antibodies (Ab3). The proportion of these two types of Ig in the serum of an Ab2-immunized animal is still a subject of investigation [3] and the study of the relationship between Abl' and Abl appears as an additionnal tool to explore the B cell repertoire involved in a specific antigenic response. The immune response against p~l y ( G l u~~, A l a~~, T y r '~) (GAT) offers a good experimental model to study these two types of questions, Different Id specificities have been characterized on the anti-GAT antibodies using a variety of anti-Id reagents .…”