Isolation and characterization of a unique C57BL B-tropic virus

A unique B-tropic virus (CS43) has been isolated from a C57BL mouse. This virus differs from those we have previously characterized and from several isolates we have obtained from normal and leukemic C57BL mice. Unlike other C57BL B-tropic viruses, CS43 does not have a p12 polypeptide homologous to that of the class II xenotropic virus, but does have a p12 homologous to that of AKR MuLV (class I). In common with the other C57BL B-tropic isolates, CS43 has an altered major core protein (p30), which competes only weakly in a class-specific assay that distinguishes ecotropic MuLV p30s from those of xenotropic origin. The data are compatible with a unique recombinational event in the generation of this isolate, which should prove useful in studying the origin of B-tropic viruses and the properties that determine N/B tropism.     

Host range conversion of murine leukemia virus resulting from recombination with endogenous virus

Summary. Ecotropic murine leukemia viruses (MuLVs) are classified into B-, N-, or NB-tropic MuLV by their host range determined by the Fv-1 gene product. B-tropic MuLV is restricted in N-type mouse cells (Fv-1^n/n) and N-tropic MuLV is restricted in B-type mouse cells (FV-1^b/b). Although forced passages in a restrictive host grant a wider host range (NB-tropism), we show here a host range conversion from B to N tropism. The conversion was most likely a result of recombination between the exogenously infected B-tropic MuLV and an endogenously expressed N-tropic MuLV in a C57BL/6 mouse cell line, YH-7. Received June 19, 1996 Accepted August 24, 1996     

Polymorphism among the major core proteins of C57BL B-tropic murine leukemia viruses

Previous studies have demonstrated a correlation between changes in the major core protein (p30) of murine leukemia viruses (MuLV) and their Fv-1-mediated host range. In this report, we have examined the p30 proteins of B-tropic viruses representing two serologically distinguishable classes. These proteins differed from each other as well as from prototype N-tropic and xenotropic viruses. This polymorphism, considered with serological typing of the adjacent p12 polypeptide, suggests that a crossover within the region coding for p30 is involved in the generation of these B-tropic viruses.     

Characterization of Fv-1 host range strains of murine retroviruses by titration and p30 protein characteristics

A standardized, direct XC plaque assay was used to determine the titration hitness patterns of Fv-1 host range murine retroviruses obtained from various laboratories. The N- or B-tropic viruses were tested on a variety of cells with Fv-1ⁿⁿ, Fv-1^bb, or Fv-1^nb genotypes, and, with one exception, a discrete two-hit pattern was obtained on cells with the restrictive genotype (i.e., N-tropic virus on BALB/c, SIM·R, and B6C3F₁ cells, and B-tropic virus on SME, SIM, and B6C3F₁ cells). The single exception to the two-hit titration effect was a strain (TOR-B) which is poorly infectious for all cells, and which does not show a clear N- or B-tropism. In addition, it was possible to convert the two-hit pattern of another B-tropic virus (OR-B) to a one-hit curve by coinfection with an XC-negative N-tropic virus. Relative to SC-1 (Fv-1^??) cells, it was possible to define three components of restriction of ecotropic virus infection of mouse cells: The first two components, the two-hit kinetics and 10^2- to 10³-fold reduction in titer on restrictive cells, are Fv-1 determined; the third a decreased infectivity on all cells relative to SC-1 cells, is independent of Fv-1 genotype.     

Naturally occurring leukemia viruses in H-2 congenic C57BL mice. III. Characterization of C-type viruses isolated from lymphomas induced by milk transmission of B-ecotropic virus

The prevalence of different host-range classes of murine leukemia virus (MuLV) was studied in C57BL mice with (V+) and without (V-) milk transmission of a naturally occurring B-tropic ecotropic MuLV. Virus isolates were studied with respect to growth properties, XC-plaque formation, antigen profiles of their envelope proteins (gp70 and P15(E)), gp70 tryptic-peptide maps, and their potential to induce lymphomas after inoculation into newborn mice. B-tropic ecotropic MuLV with the capacity to cause plaques in XC cells was isolated from almost all lymphomas of both V+ and V- sublines. The reaction patterns of these ecotropic isolates with monoclonal antibodies reactive with MuLV-env proteins and the tryptic-peptide maps of the gp70 molecule indicate that they are similar to each other and differ only slightly from the ecotropic MuLV in the spleens of young V+ animals, which is identical to the milk-transmitted virus. XC-, B-tropic dualtropic mink cell focus-inducing (MCF) viruses were isolated from the majority of different types of lymphoma (B cell, T cell, or neither B nor T cell derived), but not from the spleens or milk of young V+ or V- animals. The env proteins of the MCF isolates are highly heterogeneous, but most isolates originating from B10.AV + T-cell lymphomas share MCF-related epitopes in their gp85 envelope precursor with AKR MCF-247 virus. Most MCF viruses isolated from non-T lymphomas do not possess these epitopes. The results indicate that also in this model the generation of dualtropic MCF viruses might be important in lymphoma induction, although only some of the cloned MCF viruses show enhanced oncogenic properties in comparison with ecotropic isolates. A cloned oncogenic MCF virus induced different lymphoma types in C57BL/10 (= B10, H-2b) and B10.A (H-2a) mice, similar to what was found earlier with the milk-transmitted virus. Hence, the lymphoma-type differences are not due to differences in the B-tropic ecotropic viruses transmitted through the milk in these strains, but reflect an influence of the H-2 complex on the phenotype of the virus-induced lymphomas.     

The absence of one class of virus-induced protein in arginine-deprived cells infected with pig herpesvirus.

C57BL/6 mice responded to immunization with purified gp71 of Friend murine leukemia virus by mounting both humoral and cell-mediated responses. As measured by a number of tests, the responses were generally stronger than those obtained in BALB/c mice. However, in contrast to the BALB/c mice, immunization of C57BL/6 mice with gp71 did not result in the development of cytotoxic lymphocytes, although spleen lymphocytes were capable of undergoing blastogenesis when incubated with purified gp71. As in the BALB/c mice, the humoral response was type-specific.A unique feature of the response in gp71-immunized C57BL/6 mice was the accelerated activation of the endogenous virus as measured by the development of an immune response to its distinct envelope antigens. This resulted in the production of three distinguishable antibody populations: (1) type-specific antibodies to FLV gp71; (2) type-specific antibodies reacting with AKR gp71 (AKR virus being related to the endogenous virus of C57BL/6 mice); and (3) antibodies directed against p15 (E) which reacted both with AKR and Friend-Moloney-Rauscher viruses and are therefore considered group-specific in the murine system. The possible significance of the activation of the endogenous virus subsequent to gp71 immunization of C57BL/6 mice is discussed.     

A dominant modifier of transgene methylation is mapped by QTL analysis to mouse chromosome 13

The single-copy hepatitis B virus transgene in the E36 transgenic mouse strain undergoes methylation changes in a parent-of-origin, tissue, and strain-specific fashion. In a C57BL/6 background, the paternally transmitted transgene is methylated in 30% of cells, whereas it is methylated in more than 80% of cells in (BALB/c x C57BL/6) F1 mice. We established previously that several genetic factors were likely to contribute to the transgene methylation profile, some with demethylating and some with de novo methylating activities. Using quantitative trait loci (QTL) mapping, we have now localized one major modifier locus on chromosome 13 (Mod13), which explains a 30% increase in the methylation level of this transgene with no effect on the flanking endogenous sequences. No other QTL could be identified, except for a demethylating activity of low significance located on chromosome 12. Recombinant inbred mice containing a BALB/c allele of Mod13 were then used to show that the presence of Mod13 is sufficient to induce de novo methylation. A segregation between de novo methylation and repression of transgene expression was uncovered, suggesting that this genetic system is also useful for the identification of factors that interpret methylation patterns in the genome.     

Inherited resistance to N- and B-tropic murine leukemia viruses in vitro: evidence that congenic mouse strains SIM and SIM.R differ at the Fv-1 locus

The congenic mouse strains SIM and SIM.R, known to differ at a locus that determines susceptibility or resistance to Friend leukemia virus in vivo, were examined for their responses to murine leukemia viruses of different tropism. Responses in vitro were studied by the XC cell plaque assay method in mouse embryo cultures prepared from the 2 strains. With N-tropic viruses, SIM cells were found about 1,000-fold more sensitive than SIM.R cells; with a B-tropic virus SIM.R cells were 100- to 1000-fold more sensitive than SIM cells. The sensitivities of SIM and SIM.R cells to an NB-tropic virus were approximately equal. Similar results were obtained in vivo by the spleen focus assay method with N- and NB-tropic Friend viruses. Cells of reciprocal (SIM × SIM.R) F₁ hybrids were found resistant to both N- and B-tropic viruses.     

Effect of murine host genotype on MCF virus expression, latency, and leukemia cell type of leukemias induced by Friend murine leukemia helper virus

Leukemias induced by neonatal inoculations of several mouse strains with different strains of Friend murine leukemia helper virus (F-MuLV) were followed for time of disease onset, cytochemical analysis of predominant cell types in leukemic organs, and expression of infectious mink cell focus-inducing (MCF) viruses detected by mink cell foci or MCF-specific monoclonal antibodies. Most BALB.B and IRW mice had a rapidly appearing, severe anemia and hepatosplenomegaly consisting of erythroid cells. MCF viruses were usually isolated from enlarged spleens of IRW mice. In contrast, C57BL/10 mice had a lower incidence of disease and much slower course. Splenomegaly and lymphadenopathy with mild anemia were seen, and the predominant cell types were either myeloid (chloroleukemia) or lymphoid. MCF viruses were never isolated from this mouse strain. (C57BL/10 X IRW)F1 mice were intermediate in latency, but all mice had disease by 8 months. Myeloid, lymphoid, and some mixed leukemias with an erythroid component were observed, but in no case did we see the severe anemia or pure erythroid involvement typical of IRW and BALB.B mice. MCF viruses were, however, isolated from 22% of these mice regardless of leukemia cell type. DBA/2 mice had a disease pattern similar to the (C57BL/10 X IRW)F1 mice, and MCF viruses were isolated from three of six mice tested. Inoculation of IRW mice with the low virulence B3 strain of F-MuLV produced disease with a longer latency than F-MuLV 57, but similar cell types were transformed by both viruses. In vitro cell lines were derived from 14 mice, and most were tumorigenic in vivo. Three lines released infectious MCF virus, and three others expressed MCF-specific cell surface antigens but did not release virus. Eight lines expressed no MCF infectious virus or viral antigens. Several lines released infectious xenotropic viruses and/or expressed xenotropic MuLV cell surface antigens recognized by monoclonal antibodies reactive with xenotropic viruses. The lack of MCF expression in many primary leukemic tissues as well as in in vitro derived leukemia cell lines of C57BL/10 and (B10 X IRW)F1 mice suggested that MCF virus generation and expression may not be required for leukemogenesis in some mouse strains or in some hemopoietic lineages.     

Characterization of inducible type-C RNA viruses of mouse strains from different geographic areas

The distribution of endogenous type-C RNA viruses was studied in inbred strains of mice and some subspecies of Mus musculus of different geographical origins. The following groups of inducible viruses were characterized by their host range and immunological properties: (1) viruses indistinguishable from one of the three prototype viruses endogenous to BALB/c mice; (2) viruses coding for proteins immunologically related to different prototype endogenous viruses; (3) viruses whose p12 structural proteins were immunologically indistinguishable from that of BALB:virus-2, but whose p30 major structural proteins and envelope glycoproteins differed immunologically from those of previously characterized endogenous viruses. These findings suggest that endogenous viruses have undergone numerous genetic interactions during the process of evolution leading to inducible viruses of present day mouse strains. A class of xenotropic virus spontaneously released by NZB mice is endogenous to but not inducible from embryo cells of other previously studied mouse strains. Viruses which could not be distinguished from the NZB xenotropic virus by host range analysis or radioimmunological techniques were chemically inducible from embryo cells of several mouse strains originating in Asia and Europe. These results indicate that the biological regulatory mechanisms that affect expression of this virus have evolved differently in such strains from control mechanisms that developed in standard inbred strains.     

Differential susceptibility to acute Trypanosoma cruzi infection in BALB/c and C57BL/6 mice is not associated with a distinct parasite load but cytokine abnormalities

Inoculation of Trypanosoma cruzi, Tulahuén strain, into C57BL/6 and BALB/c mice led to an acute infection characterized by marked parasitaemia, myocardial inflammation and thymocyte depletion. While C57BL/6 mice showed a progressive and lethal disease, BALB/c mice partly recovered. To characterize these murine models more effectively, we studied the parasite burden, serum levels of major infection outcome-related cytokines, the in vitro features of T. cruzi infection in peritoneal macrophages and the immunophenotype of thymic cells. The greater disease severity of T. cruzi-infected C57BL/6 mice was not linked to an increased parasite load, as parasitaemia, myocardial parasite nests and amastigote counts in peritoneal macrophages were not different from those in BALB/c mice. Cortical thymocyte loss was accompanied by the presence of apoptotic bodies and fragmented nuclear DNA, whereas fluorocytometric analysis at 17 days postinfection (p.i.) revealed a more pronounced loss of CD4+ CD8+ cells in C57BL/6 mice. This group displayed higher levels of TNF-alpha on days 14 and 21 p.i., in the presence of lower IL-1beta and IL-10 concentrations by days 14 and 21, and days 7 and 14 p.i., respectively. Day-21 evaluation showed higher concentrations of nitrate and TNF-alpha soluble receptors in C57BL/6 mice with no differences in IFN-gamma levels, with respect to the BALB/c group. Increased morbidity of C57BL/6 T. cruzi-infected mice does not seem to result from an aggravated infection but from an unbalanced relationship between pro- and anti-inflammatory mediators.     

Immune responses to Yersinia enterocolitica in susceptible BALB/c and resistant C57BL/6 mice: an essential role for gamma interferon.

Susceptibility of mice to infection with Yersinia enterocolitica has been shown to be related to neither the Ity locus encoding for resistance to Salmonella typhimurium and other pathogens nor the H-2 locus. Recent studies in our laboratory have demonstrated that T-cell-mediated immune responses are required for overcoming primary Yersinia infection. In the present study, we investigated the course of infection with Y. enterocolitica and the resulting immune responses in Yersinia-susceptible BALB/c and Yersinia-resistant C57BL/6 mice. In the early phase of infection, the clearance of the pathogen was comparable in both strains of mice, suggesting similar mechanisms of innate resistance. Splenic T cells from Yersinia-infected C57BL/6 mice exhibited marked proliferative responses and produced gamma interferon (IFN-gamma) upon exposure to heat-killed yersiniae. By contrast, the Yersinia-specific T-cell response in BALB/c mice was weak, and IFN-gamma production could not be detected before day 21 postinfection. T cells isolated from C57BL/6 mice 7 days after infection mediated immunity to Y. enterocolitica but those from BALB/c mice did not, while at 21 days postinfection T cells from both strains mediated protection. Neutralization of IFN-gamma abrogated resistance to yersiniae in C57BL/6 mice but to a far smaller extent in BALB/c mice. Administration of recombinant IFN-gamma or anti-interleukin-4 antibodies rendered BALB/c mice resistant to yersiniae, whereas this treatment did not significantly affect the course of the infection in C57BL/6 mice. These results indicate that the cellular immune response, in particular the production of IFN-gamma by Yersinia-specific T cells, is associated with resistance of mice to Y. enterocolitica.     

Irf4 is a positional and functional candidate gene for the control of serum IgM levels in the mouse

Natural IgM are involved in numerous immunological functions but the genetic factors that control the homeostasis of its secretion and upholding remain unknown. Prompted by the finding that C57BL/6 mice had significantly lower serum levels of IgM when compared with BALB/c mice, we performed a genome-wide screen and found that the level of serum IgM was controlled by a QTL on chromosome 13 reaching the highest level of association at marker D13Mit266 (LOD score=3.54). This locus was named IgMSC1 and covered a region encompassing the interferon-regulatory factor 4 gene (Irf4). The number of splenic mature B cells in C57BL/6 did not differ from BALB/c mice but we found that low serum levels of IgM in C57BL/6 mice correlated with lower frequency of IgM-secreting cells in the spleen and in the peritoneal cavity. These results suggested that C57BL/6 mice have lower efficiency in late B-cell maturation, a process that is highly impaired in Irf4 knockout mice. In fact, we also found reduced Irf4 gene expression in B cells of C57BL/6 mice. Thus, we propose Irf4 as a candidate for the IgMSC1 locus, which controls IgM homeostatic levels at the level of B-cell terminal differentiation.     

Identification of the major structural proteins of two BALB/c myeloma C-type viruses.

We have analyzed the structural proteins of the C-type viruses produced by cloned lines of the MOPC-21 and FLOPC-1 BALB/c murine myelomas. These viruses, designated MO₂₁-MuMAV and FL₁-MuMAV, respectively, have been shown to be composed of proteins with approximate molecular weights of 90,000, 75,000, 17,000, 12,000, and 10,000 daltons. In addition, these viruses contain a “doublet” with molecular weights of approximately 30,000 daltons. The 90,000- and 75,000-dalton proteins are glycosylated. The p30 “doublet” as well as the NB-ecotropism is retained upon subsequent virus cloning in SC-1 cells, passage through NIH-3T3, or BALB/3T3 cells, or microtiter plate cloning of SC-1 cells replicating cloned virus. The two cloned MuMAVs appeared to possess the p30 proteins of the endogenous BALB/c N- and B-tropic viruses described by Hopkins and co-workers (N. Hopkins, J. Schindler, and R. Hynes, 1977, J. Virol.21, 309–318; J. Schindler, R. Hynes, and N. Hopkins, 1977, J. Virol. 23, 700–707). Subsequent tryptic peptide analysis of the myeloma virus p30 proteins demonstrated that these proteins share extensive homology with, but not identity to, the two prototype BALB/c ecotropic endogenous virus p30 proteins.     

Structural heterogeneity in p30 molecules of type C viruses

Tryptic digests of p30 proteins from mouse type C viruses were subjected to cation-exchange chromatography. Structural heterogeneity of p30 molecules was seen in two specific areas of the peptide elution profiles. These hypervariable regions of p30 proteins were used to discriminate representative ecotropic (N- and B-tropic), xenotropic (alpha and beta) and amphotropic viruses.     

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

Two monoclonal anti-idiotypic antibodies (HP-Id20 and HP-Id22) recognizing two different public idiotopes expressed in the anti-poly(Glu60,Ala30,Tyr10) (GAT) response were used to immunize BALB/c and C57BL/6 mice. From these animals hybridomas were isolated. From BALB/c and C57BL/6 mice eight and seven monoclonal antibodies were characterized, respectively. The reagents were classified according to the expression of the public idiotypic specificity p.GAT (recognized by a rabbit antiserum). The anti-GAT activity and the expression of the various idiotopes characterized on anti-GAT polyclonal and monoclonal antibodies were also studied. Most of the reagents are Ab1'-type of antibody resembling anti-GAT antibodies. One anti-anti-idiotypic monoclonal antibody (Ab3) was also isolated from BALB/c mice. This suggests that in this experimental model the repertoire induced after HP-Id immunization and antigen stimulation is comparable. The idiotypic analysis of a large number of anti-GAT and of Ab1' monoclonal antibodies suggests that only two public idiotopes are involved in the anti-GAT response.     

Differences in expression of toll-like receptors and their reactivities in dendritic cells in BALB/c and C57BL/6 mice

We have previously reported that differences in early production of interleukin 12 (IL-12) by dendritic cells (DC) underlies the difference between the susceptibilities to Listeria monocytogenes of C57BL/6 and BALB/c mice. To elucidate mechanisms for the different abilities of DC to produce cytokine in C57BL/6 and BALB/c mice, we examined Toll-like receptor (TLR) expression by DC and their responses in vitro to known microbial ligands for TLRs. We found that DC isolated from the spleens of naive C57BL/6 mice preferentially expressed TLR9 mRNA, whereas DC from naive BALB/c mice strongly expressed TLR2, -4, -5, and -6 mRNAs. C57BL/6 DC produced a higher level of IL-12p40 in response to the ligands for TLR4 (lipopolysaccharide), TLR2 (lipoprotein), and TLR9 (CpG), whereas BALB/c DC responded to these ligands by producing a larger amount of monocyte chemoattractant protein 1. C57BL/6 DC expressed higher levels of CD40 and Stat4 than BALB/c DC did, suggesting that naive C57BL/6 mice contained more-mature subsets of DC than naive BALB/c mice. Differences in reactivities of DC to microbial molecules through TLRs may be associated with susceptibility and resistance to Listeria infection in BALB/c and C57BL/6 mice.     

In vitro and in vivo expression analysis of the interferon-inducible 203 gene.

The interferon (IFN)-inducible protein family 200 is encoded by structurally related genes located on mouse chromosome 1. The encoded proteins so far characterized and designated p202, p204, and pD3 contain at least one copy of a conserved 200 amino acid domain in addition to other regions that are different or missing among the various family members. We have recently characterized a cDNA clone (203 cDNA) encoding a 408 amino acid protein bearing structural similarities to p202 and p204. Here, we report its pattern of expression in vitro and in vivo. In vitro, the mRNA and protein encoded by the 203 gene were increased by IFN-alpha in several cell lines of different histologic origin. By contrast, no significant induction was observed in vivo in mice from C57BL/6 and BALB/c strains even after treatment with the IFN-inducer poly rI:rC. In addition, the constitutive expression of 203 gene was restricted to some myeloid and lymphoid tissues, namely, thymus, bone marrow, and spleen. Comparison of the expression pattern of the 203 and 202 genes in three mouse strains revealed that they exhibit a differential inducibility by IFN and a reciprocal expression pattern. The 203 mRNA was constitutively expressed in C57BL/6 and BALB/c mice and undetectable in the spleen of DBA/2 mice. The 202 mRNA was strongly induced by poly rI:rC in the spleen of DBA/2 and BALB/c mice but absent in C57BL/6 mice. Southern analysis revealed a restriction fragment length polymorphism in the 203 locus. Taken as a whole, these results demonstrate a remarkable difference in the in vivo IFN responsiveness of two members belonging to the same gene family with a similar degree of IFN inducibility in vitro. Moreover, the reciprocal expression pattern in C57BL/6 and DBA/2 mice could mean that p203 and p202 play the same role in a mouse strain in which only one of them is expressed.     

Regulation by endogenous interferon of virus-induced cytokine gene expression in mouse macrophages.

In macrophages from inbred mice the magnitude of the interferon (IFN) response to Newcastle disease virus (NDV) infection is under genetic control of the locus If-1, with C57BL/6 carrying the 'high-producer' allele If-1h whereas BALB/c have the 'low-producer' allele If-1l. The IFN produced consists of 90% IFN-beta and there are 10-fold differences between macrophages from If-1h and If-1l mice. Recently, we observed that interleukin-6 (IL-6) is coinduced by NDV in macrophages and seems to be under the same genetic control. Noninduced macrophages have been shown to secrete low amounts of antiviral activity endogenously when cultured in the presence of the macrophage-colony-stimulating factor (M-CSF). Here, we report that the amount of this endogenous IFN varies between macrophages from different mouse strains. Macrophages from BALB/c were found to secrete 5-10 times more endogenous IFN compared to C57BL/6. The antiviral activity could be identified as IFN-beta. Interestingly, we observed that endogenous IFN specifically down-regulates NDV-induced IFN and IL-6 production. Preculture of BALB/c macrophages in M-CSF plus anti-IFN-beta to neutralize the biological effects of the endogenous IFN provoked a 30- to 50-fold increase in NDV-induced cytokine production, resulting in a nearly complete abrogation of the genetically determined difference since the same treatment only caused a 6-fold increase in C57BL/6 macrophages following NDV infection. This increase in cytokine gene expression was specific for NDV and marked by a strong additional activation of IFN-alpha genes. Addition of mouse recombinant IFN-alpha 4 to anti-IFN-beta-treated macrophages for 18 h prior to NDV infection down-regulated again IFN gene expression and reestablished the genetic differences between macrophages from If-1h and If-1l mice.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of genetic factors associated with the immunoglobulin heavy chain locus on the development of benign monoclonal gammapathy in ageing IgH-congenic mice.

The role of genetic factors associated with the immunoglobulin heavy chain locus (Igh) in the development of benign monoclonal gammapathy (BMG), a benign B-cell proliferative disorder, was investigated in six Igh congenic mouse strains during ageing. The strains used had a C57BL or BALB background: C57BL/6, BALB.Igb and CB-20 carrying the C57BL Igh (Ighb allotype), BALB/c and C57BL/6.Iga carrying the BALB/c Igh (Igha allotype) and BAB-14, that is of BALB/c origin with the exception of the constant part of the Igh, which is of C57BL origin. The frequency of homogeneous immunoglobulins (H-Ig), both single and multiple, was the highest in C57BL/6 mice, followed by C57BL/6.Iga. The frequencies of H-Ig in BALB.Igb and CB-20 mice were higher than those of BALB/c and BAB-14, although somewhat lower than in C57BL/6.Iga mice. Multiple H-Ig were found especially in the sera of C57BL/6 mice. Categorization of the monoclonal gammapathies (MG) on the basis of their origin showed a single transient monoclonal B-cell proliferation in 0-8% of the mice of all strains. Persistent, non-progressive MG, presumably BMG, were detected in 64% of C57BL/6, 30% of C57BL/6.Iga, 22% of BALB.Igb, 17% of CB-20, 13% of BAB-14 and 6% of BALB/c mice. Multiple myeloma or Waldenström-like B-cell lymphoma were found to be responsible for 2-4% of the paraproteinemias in all strains. The remaining H-Ig, varying from 11% of the C57BL/6 to 70% of the BAB-14 mice, could not be evaluated in time. The most frequent isotypes of the BMG within C57BL/6 and C57BL/6.Iga were IgG2a and IgG2b, respectively; IgM was the most frequent isotype within the four BALB congenic strains. The immunoglobulin heavy chain allotypes under investigation appeared to be only partly related to the onset, occurrence, multiplicity and persistence of the BMG developing in these Igh congenic C57BL and BALB strains during ageing. The immunoglobulin heavy chain allotypes, however, were not related to the major isotype of the BMG. The results obtained in CB-20 and BALB.Igb on the one hand, and in BAB-14 on the other hand, may suggest a role for the variable part of the Igh in the development of BMG. Since no absolute influence could be ascribed to the Igh, we assume that primarily other genetic sequences regulating proliferative B-cell functions account for the pathogenesis of BMG.