Polyclonal IgM obtained by fractionation of a BALB/c serum pool was used as the immunogen for C57BL/6 mice. Draining lymph nodes from selected animals donated cells for fusion with myeloma Sp-2/0. Fifteen hybridomas were productive, and one (RS-3.1) was cloned and the affinity-purified product analyzed for its reactivity pattern by MOPC 104E-enzyme-linked immunosorbent assay inhibition. Among five BALB/c myelomas only TEPC 183 (IgM) was active, not those belonging to other Ig classes. Among normal sera from 8 mouse strains only those of BALB/c, DBA/2J and CBA/J showed inhibition. The recombinant strain BALB-Igh-Va/Igh-Cb did not react, which shows that its C mu stems from parental strain C57BL/6, and that therefore the recombination event had occurred 5' of this gene.
Eight isogeneic anti-idiotypic hybridomas were raised against BALB/c myeloma protein MOPC 104E and one against J558. Both myelomas react specifically with the alpha(1----3) glucosidic linkage of dextran B1355 fraction S (Dex). Six anti-MOPC 104E proteins were IgG1, one was IgG2b and one IgM. The anti-J558 protein was IgG1. Competitive interactions of the anti-idiotopes and antigen with anti-Dex proteins were measured. Dex itself was effective, but also an alpha(1----3) glucosidic heptasaccharide (N7-CHO). In order to assess the anti-idiotope specificity of hybridoma proteins, three anti-Dex molecules were used: MOPC 104E, J558 and hybridoma protein Hdex14. These differed from each other in VH amino acid positions 54-55, or 100-101, respectively. By their serological reaction pattern our anti-idiotope proteins could be divided into 3 groups: cross-reactive, partially cross-reactive and strictly specific for the immunogen. The latter ones were in the majority, and were called "private", in contrast to the cross-reactive "public" anti-idiotopes. The serological pattern was followed, in general, by the mouse-to-mouse distribution of idiotopes in physiological anti-Dex sera. Public idiotopes were closely correlated in their expression with anti-Dex activity. "Private" idiotopes showed no correlation, and displayed a characteristically high degree of fluctuation from mouse to mouse. Among the different mouse strains that were compared with respect to idiotope expression in anti-Dex sera, two stand out: C57BL-Igha, which carries chromosome 12 of BALB/c, (as selected through allotype) on the C57BL/6 genome, and BALB-Ighb, dex+, a recombinant in chromosome 12 linking the dex+ trait from BALB/c to the CH allotype from C57BL/6. The latter strain expressed significantly more of the private idiotopes than the former. This observation is discussed in terms of the position effect of classical genetics and network concepts.
From a panel of isogeneic monoclonal anti-idiotope antibodies several were used as agents in neonatal idiotope suppression. They differed from one another in isotype, and in idiotope specificity, as described in the preceding report (Eur. J. Immunol. 1987. 17: 255). In their effects they were compared with respect to the following variables: minimum dose required for suppression; duration of suppression, and its relationship to the dose applied neonatally; half-life of anti-idiotope in the immune system of the young mice; specificity of suppression as achieved by a given anti-idiotope: in how far does it affect idiotopes defined by alternate anti-idiotopes? The following results were obtained: the minimum effective dose varied widely between anti-idiotopes. One, belonging to the IgM class, was completely ineffective; others varied from approximately 10 micrograms/mouse, required for complete suppression, to approximately 100 micrograms/mouse. The dose-response characteristic was independent of whether the state of suppression was tested (by immunization against alpha(1----3)dextran) 26 days or 70 days after neonatal anti-idiotope treatment. We take this as an indication that the anti-idiotope effect occurs during an early postnatal period. There appeared to be a relationship between the rate of decay of anti-idiotope in the system and the dose required for complete suppression: the faster the decay, the more is needed initially. The persistence of effective molecules in the animals appears to depend on their isotype (as has been noted by others before): IgM decays fastest, and was ineffective in our experiments; IgG1 stays longest, and the smallest dose was required for suppression. IgG2b was intermediate. The specificity of neonatal suppression was clearly correlated with the serological specificity of the anti-idiotope monoclonal antibodies, as well as with the representation of the corresponding idiotopes in physiological anti-dextran sera, as described in the preceding report: private anti-idiotopes suppressed their counterpart idiotopes only, while the public anti-idiotope suppressed all other idiotopes in concert.
The aim of our investigations was to discriminate unspecific age-depending function of the tonsil. In an in-vitro system of tonsillar lymphocytes we could show, that there are differences in the DNA-, RNA- and Proteinbiosynthese between children and adults. The children organ is more active. It could be shown that the observed differences are only age depending and not influenced by the clinical state of the organ.
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