Comparative serum metabolomics between SCID mice and BALB/c mice with or without Schistosoma japonicum infection: Clues to the abnormal growth and development of schistosome in SCID mice.

The small blood flukes of genus Schistosoma, which cause one of the most prevalent and serious parasitic zoonosis schistosomiasis, are dependent on immune-related factors of their mammalian host to facilitate their growth and development, and the formation of granulomatous pathology caused by eggs deposited in host's liver and intestinal wall. Schistosome development is hampered in the mice lacking just T cells, and is even more heavily retarded in the severe combined immunodeficient (SCID) mice lacking both T and B lymphocytes. Nevertheless, it's still not clear about the underlying regulatory molecular mechanisms of schistosome growth and development by host's immune system. This study, therefore, detected and compared the serum metabolic profiles between the immunodeficient mice and immunocompetent mice (SCID mice vs. BALB/c mice) before and after S. japonicum infection (on the thirty-fifth day post infection using liquid chromatography-mass spectrometry (LC-MS). Totally, 705 ion features in electrospray ionization in positive-ion mode (ESI+) and 242 ion features in ESI- mode were identified, respectively. First, distinct serum metabolic profiles were identified between SCID mice and BALB/c mice without S. japonicum worms infection. Second, uniquely perturbed serum metabolites and their enriched pathways were also obtained between SCID mice and BALB/c mice after S. japonicum infection, which included differential metabolites due to both species differences and differential responses to S. japonicum infection. The metabolic pathways analysis revealed that arachidonic acid metabolism, biosynthesis of unsaturated fatty acids, linoleic acid metabolism, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, alpha-linolenic acid metabolism, glycerophospholipid metabolism, sphingolipid metabolism and purine metabolism were enriched based on the differential serum metabolites between SCID mice and BALB/c mice after S. japonicum infection, which was addressed to be related to the retarded growth and development of S. japonicum in SCID mice. These findings provide new clues to the underlying molecular events of host's systemic metabolic changes on the growth and development of S. japonicum worms, and also provide quite promising candidates for exploitation of drugs or vaccines against schistosome and schistosomiasis.

Sociability and brain development in BALB/cJ and C57BL/6J mice.

Sociability--the tendency to seek social interaction--propels the development of social cognition and social skills, but is disrupted in autism spectrum disorders (ASD). BALB/cJ and C57BL/6J inbred mouse strains are useful models of low and high levels of juvenile sociability, respectively, but the neurobiological and developmental factors that account for the strains' contrasting sociability levels are largely unknown. We hypothesized that BALB/cJ mice would show increasing sociability with age but that C57BL/6J mice would show high sociability throughout development. We also hypothesized that littermates would resemble one another in sociability more than non-littermates. Finally, we hypothesized that low sociability would be associated with low corpus callosum size and increased brain size in BALB/cJ mice. Separate cohorts of C57BL/6J and BALB/cJ mice were tested for sociability at 19-, 23-, 31-, 42-, or 70-days-of-age, and brain weights and mid-sagittal corpus callosum area were measured. BALB/cJ sociability increased with age, and a strain by age interaction in sociability between 31 and 42 days of age suggested strong effects of puberty on sociability development. Sociability scores clustered according to litter membership in both strains, and perinatal litter size and sex ratio were identified as factors that contributed to this clustering in C57BL/6J, but not BALB/cJ, litters. There was no association between corpus callosum size and sociability, but smaller brains were associated with lower sociability in BALB/cJ mice. The associations reported here will provide directions for future mechanistic studies of sociability development.

Study of embryotoxicity of mentha piperita L. during organogenesis in balb/c mice / Estudio de la embriotoxicidad de mentha piperita L. durante la organogénesis en ratones balb/c

Mentha piperita (Labiatae), commonly known as peppermint is a native Iranian herb which is used in folk medicine for various purposes. This study was carried out to reveal the teratogenic effect of Mentha piperita on mice fetuses. In this experimental study, pregnant Balb/c mice divided to four groups. Case group received 600 (treatment I) and 1200 (treatment II) mg/kg/day the hydroalcoholic extract of Mentha piperita during 6-15 of gestational days and one control group received normal saline during GD6-GD15 by gavages and other control group did not receive any matter during 6-15 of gestational days. Mice sacrificed at GD18 and embryos were collected. Macroscopic observation was done by stereomicroscope. 20 fetuses of each group were stained by Alizarin red-S and Alcian blue staining method. The Mean weight of fetuses decreased in treatment groups rather than control (P<0.05) but CRL there was no significant difference between treatments and controls groups. In the treatment I (600 mg/kg/day) and treatment II (1200 mg/kg/day), normal saline and control group, no gross congenital malformations were observed in fetuses. Treated fetuses also had no delayed bone ossification as determined by Alizarin red-S and Alcian blue staining method. This study showed that the hydroalcoholic extract of Mentha piperita (600 and 1200 mg/kg/day) has no teratogenic effect in mice fetuses if used continuously during embryonic period.

Mentha piperita (Labiatae), comúnmente conocida como menta, es una hierba nativa de Irán, que se utiliza en la medicina tradicional para diversos fines. Este estudio fue realizado para descubrir el efecto teratogénico de la Mentha piperita en fetos de ratones. Los ratones Balb/c preñadas fueron divididas en cuatro grupos. Los grupos recibieron 600 (tratamiento I) y 1200 (tratamiento II) mg/kg/día del extracto hidroalcohólico de Mentha piperita durante los días 6-15 de gestación (DG), mientras que un grupo control recibió solución salina normal durante los DG 6-15 vía oral y otro grupo control sano no recibió substancia durante los DG 6-15. Los ratones fueron sacrificados el DG 18, recolectando los fetos. Se realizó la observación macroscópica mediante un estereomicroscopio. 20 fetos de cada grupo se tiñeron por el método de rojo de alizarina-S y azul de Alcián. La media de peso de los fetos disminuyó más en los grupos de tratamientos que los controles (p <0,05), pero CRL no presentó diferencias significativas entre los tratamientos y los grupos control. En los fetos del grupos tratamiento I (600 mg/kg/día), tratamiento II (1200 mg/kg/día), solución salina normal y control no se observó ninguna malformación congénita grave. Los fetos tratados tampoco tuvieron osificación ósea retrasada según lo determinado por el método de rojo de alizarina-S y azul de Alcián. Este estudio mostró que el extracto hidroalcohólico de Mentha piperita (600 y 1200 mg/kg/día) no tiene efectos teratogénicos en fetos de ratones al ser utilizado continuamente durante el período embrionario.

Increased juvenile and adult body weights in BALB/cByJ mice reared in a communal nest.

Both wild and laboratory mice and rats preferentially rear their young in communal nests and indiscriminately nurse any of the young within the nest. In this study, BALBc/ByJ mice reared under communal nesting (CN) conditions (3 dams and their litters sharing a common nest) were compared with BALBc/ByJ mice raised in single (one dam with her litter) nests (SN) in body weight from birth into adulthood; food and water intake and body composition were compared between adult mice. Compared with SN female mice, female CN mice (measured only until weaning) exhibited significantly higher body weights at postnatal days 11 and 25. Male CN mice were significantly heavier than were male SN mice at postnatal day 25 and at 20, 26, and 30 wk of age. There were no differences between adult male mice from CN and SN groups in 48-h food and water intake or body composition (total lean:total fat ratio; measured by quantitative MRI). In conclusion, BALB/cByJ mice reared under communal nesting conditions showed more robust juvenile growth rates than did mice raised with a single dam and litter per cage. In addition, body weights of male CN mice remained higher than male SN mice into adulthood.

Paternal transmission of complex phenotypes in inbred mice.

BACKGROUND: Inbred mice are genetically identical but nonetheless demonstrate substantial variability in complex behaviors such as activity levels in a novel environment. This variability has been associated with levels of parental care experienced early in development. Although maternal effects have been reported in biparental and uniparental strains, there have been no investigations of paternal effects in non-biparental strains in which offspring are reared exclusively by mothers. METHODS: In the uniparental inbred Balb/cJ mouse strain, we examined the relationship of paternal open-field activity to the activity of both male and female offspring in the open-field. Potential mediators of paternal transmission of behavior were examined, including maternal care, growth parameters, litter characteristics, and time the father was present with the pregnant mother prenatally. RESULTS: An association of paternal open-field activity with the open-field activity of female but not male offspring was found. Variation in maternal postnatal care was associated with female but not male offspring activity in the open-field but did not mediate paternal effects on offspring behavior. Paternal effects on offspring growth parameters were present, but these effects also did not mediate paternal effects on behavior. CONCLUSIONS: Paternal transmission of complex traits in genetically identical mice reared only by mothers suggests a nongenetic mechanism of inheritance potentially mediated by epigenetic factors. The exclusion of multiple mediators of paternal effects on offspring suggests the possibility of germline paternal inheritance via sperm of complex phenotypes in inbred mice. Future studies are required to examine these interesting possibilities.

Teratogenic effect of lithium carbonate in early development of BALB/c mouse.

Lithium carbonate is used as a standard treatment for manic depression. While researchers have investigated the teratogenic effects of lithium carbonate on embryos of various animals in later stages of development, very limited work has been done on the probability of effects at early stages of development. In this study, the teratogenic effect of lithium carbonate was investigated at earlier preimplantation through implantation stages of development of Balb/C mouse embryos. A therapeutic dose (i.e., 300 mg/kg b.w.) was injected into mice intraperitoneally on days 3.5, 4.5, 5.5, and 6.5 of pregnancy. Then, on day 15.5 of gestation, embryos were collected from the pregnant animals. Among the embryos, 71.7% were healthy, 10.7% resorbed, 3.1% showed lordosis, 8.1% were underdeveloped and 8.4% had eye malformations. Significant increases (P < 0.05) in the number of hepatic megakaryocytes and nucleated red cells were also observed among experimental embryos. Analysis of maternal serum proteins prepared from dissected animals showed a significant increase or decrease (P < 0.05) in the levels of serum proteins albumin, alpha2 globulin, beta globulin, and gamma globulin. This research on early developmental stages suggests that pregnant mothers need to be aware of possible teratogenic effects at early stages of pregnancy, although it has been thought that the egg envelope can prevent teratogens from entering. In this case, mothers may need to stop lithium carbonate treatment before they make a decision to become pregnant.

Bruch's membrane of the brachymorphic mouse.

Bruch's membrane exists between the retinal pigment epithelium and the choriocapillary endothelium. Its structure is very complicated, having five sublayers containing basement membranes of retinal pigment epithelium and choriocapillary endothelium, outer and inner collagenous layers, and a central elastic layer. In the development of Bruch's membrane in normal mice, both basement membranes are created first. Secondarily, collagen fibers are accumulated in the space between these basement membranes and then form a collagenous layer. Finally, the elastic layer elaborated in the collagenous layer separates this into outer and inner collagenous layers. Brachymorphic mice have a disorder in the sulfation pathway, resulting in undersulfation. Consequently, in Bruch's membrane of brachymorphic mice, the expression of decorin, a small proteoglycan containing chondroitin sulfate and an indispensable component in collagen assembly, is at a very low level. It is clear that hypoplasia of the collagenous layer in Bruch's membrane of brachymorphic mice induces a disorder in the following formation of the elastic layer. These findings suggest that the formation of the collagenous layer, regulated with acidic glycoconjugates such as decorin, is important in the development of Bruch's membrane.

Marked differences in olfactory sensitivity and apparent speed of forebrain neuroblast migration in three inbred strains of mice.

In the adult forebrain, new neuroblasts constantly migrate from the subventricular zone along the rostral migratory stream to the olfactory bulb, where many become neurons. It is unclear whether this process is different in commonly used mouse strains and whether it is related to olfactory function. Adult male BALB/c, C57BL/6, and 129/S1 (formerly 129SV) mice were tested for olfactory sensitivity plus discrimination, using male mouse urine from the two other strains. BALB/c mice had the greatest olfactory sensitivity, followed by 129/S1, and C57BL/6 mice, by an order of magnitude each. Newly formed cells were pulse-labeled for 3 h with i.p. 5-bromo-2'-deoxyuridine (BrdU) injections and the animals analyzed 24 h later. In 129/S1 mice, a greater proportion of neuroblasts were present closer to the olfactory bulb than in BALB/c mice, followed by C57BL/6 mice. The total number of BrdU-labeled cells did not differ, suggesting differences in migration and not proliferation. The impaired olfactory function in C57BL/6 mice might be caused by the reduced number of neuroblasts that reach the olfactory bulbs. However, olfactory function in BALB/c and 129/S1 mice did not correlate with their putative migration speed, suggesting a more complex nature of cellular processes that contribute to olfactory function. These results caution against comparing studies of olfactory function or neural precursors that use different strains of mice, and question the use of C57BL/6 mice as a "normal" strain or as transgenic background. Perhaps more importantly, the results point to an opportunity to identify genes that regulate olfactory function and neuroblast behavior.

Expression of specific glycoconjugates in both primary and secondary olfactory pathways in BALB/C mice.

Binding of cell surface carbohydrates to their receptors specifically promotes axon growth and synaptogenesis in select regions of the developing nervous system. In some cases these interactions depend upon cell-cell adhesion mediated by the same glycoconjugates present on the surface of apposing cells or their processes. We have previously shown that the plant lectin Dolichos biflorus agglutinin (DBA) binds to a subpopulation of mouse primary olfactory neurons whose axons selectively fasciculate prior to terminating in the olfactory bulb. In the present study, we investigated whether these glycoconjugates were also expressed by postsynaptic olfactory neurons specifically within the olfactory pathway. We show here for the first time that DBA ligands were expressed both by a subset of primary olfactory neurons as well as by the postsynaptic mitral/tufted cells in BALB/C mice. These glycoconjugates were first detected on mitral/tufted cell axons during the early postnatal period, at a time when there is considerable synaptogenesis and synaptic remodelling in the primary olfactory cortex. This is one of the few examples of the selective expression of molecules in contiguous axon tracts in the mammalian nervous system. These results suggest that glycoconjugates recognized by DBA may have a specific role in the formation and maintenance of neural connections within a select functional pathway in the brain.

Aspen wood-wool is preferred as a resting place, but does not affect intracage fighting of male BALB/c and C57BL/6J mice.

Aspen wood-wool, provided as nesting material, was evaluated as a possible improvement of cage environment for 10-14-week-old inbred male mice maintained in groups of six (BALB/c n = 72 and C57BL/6J n = 36). The daily behaviour of mice was video recorded and their body weight, food consumption, weights of some organs and serum corticosterone concentrations were measured. Aggressive interactions between cage mates and against a strange intruder as well as the number of wounds on the back of the animals was monitored in order to evaluate the effect of nesting material on intermale aggression. Nesting material did not affect the daily active/passive behaviour patterns of mice, although animals clearly preferred it as a resting place. BALB/c mice given nesting material showed less weight gain and smaller brown adipose tissue weights than animals without nesting material. The other characteristics measured were not affected by the presence of nesting material in either strain. The presence of nesting material had no effect on fighting in cages. C57BL/6J mice were more aggressive than BALB/c mice according to the number of wounded animals in a cage. Wounded BALB/c mice had enlarged spleens and decreased epididymal adipose tissue weights. In conclusion, the nesting material used in this study did not adversely affect the animals. On the other hand, the material was clearly preferred to conventional bedding as a resting place. These findings suggest that nesting material may improve the cage environment of laboratory mice. Furthermore, there was an indication of strain differences in aggressive behaviour. It could be suggested that C57BL/6J mice are less tolerant towards intruders and housing six mice per cage is not suitable for this strain.

Senescent BALB/c mice exhibit decreased expression of lambda5 surrogate light chains and reduced development within the pre-B cell compartment.

Although senescent BALB/c mice (approximately 2 years old) have reduced numbers of small pre-B cells, early pre-B cells (CD43+CD25+B220+) are present in comparable numbers within the bone marrow of both young (3-6-month-old) and senescent BALB/c mice. The transition of CD43+ pre-B cells to the CD43- pre-B cell compartments is dependent on proliferation and clonal maturation dictated by the pre-B cell receptor (mu/lambda5/VpreB). In vivo, senescent CD43+B220+ pro-B/early pre-B cells demonstrated reduction of lambda5 mRNA, by RT-PCR analysis, and of both surface and cytoplasmic lambda5 protein. Decreased lambda5 protein expression was also seen among pro-B/pre-B cells derived from senescent bone marrow after stimulation in vitro with IL-7. We propose that diminished expression of the lambda5 surrogate light chain results in decreased pre-B cell receptor formation and contributes to reduced recruitment of nascent CD43+ pre-B cells into the CD43- large and small pre-B cell compartments.

Expression and distribution of the receptors for coxsackievirus B3 during fetal development of the Balb/c mouse and of their brain cells in culture.

This study was designed mainly to determine the relationships between the expression and distribution of the cellular receptor proteins for coxsackievirus B3 (CVB3) and susceptibility of mouse brain cells during fetal development of Balb/c mice. Immunoblot analysis of fetal extracts demonstrated that the CVB3 receptor proteins were first expressed at day 14 of the fetal stage, and that maximal expression of the cellular receptor occurred at near term or newborn stage. Results also suggested that newborn mouse brain tissue expressed much larger quantities of viral receptor proteins, compared to other tissues. In vitro studies showed that both mouse neurons and astrocytes could be infected by two CVB3 strains, pantropic CVB3 Nancy strain (CVB3N) and myocardiotropic CVB3 Woodruff strain (CVB3W). CVB3N, however, replicated and grew to high titer in primary astrocyte cultures and in primary neuron cultures, whereas, primary astrocyte cultures were relatively resistant to CVB3W. Virus binding assays revealed that CVB3N bound faster and in greater amounts to mouse brain cells than CVBW. These two virus strains, however, were found to share the same receptor specificity by virus competition assays. The number of virus binding sites for CVB3 on newborn mouse brain cells was approximately 1.8 x 10(4) per cell. The data suggested that preferential expression of the cellular receptors on newborn mouse brain cells may be related to their high susceptibilities to CVB3 infection.

Recombination of epithelial root sheath and dental papilla cells in vitro

Hertwing's epithelial root sheath (HERS) cells were isolated and recombined with ectomesenchymal cells in vitro utilizing extracellular matrix components as substrate. After 14 days in culture, HERS cells were differnetiated and exhibited a stratified organization. These features resembled those observed in vivo as epithelial rests of Malassez. A mineralization process was also present in HERS cells, in which calcium salts were deposited. This mineralization was correlated with the strong immunoexpression of osteopontin by HERS. The results obtained add support to the possible role of HERS in the secretion of Hypocalcified material on the root during early cementogenesis

Age-related polymorphism of thymus subpopulations in inbred mice.

There are striking age-related changes in the demography of thymus lymphocytes, i.e. in thymus-cell subpopulations of BALB/c and SJL mice; these changes occur in the proportion of cells, identified by various markers, and by the membrane density of these markers. The thymuses of both strains undergo an age-related increase in the proportion of CD4+ CD8- cells and decrease in CD4+ CD8+ cells. Age-related changes in cells that are Pgp-1+ also show marked strain differences: Pgp-1+ cells increase in SJL, but not in BALB/c thymuses. In both strains, cells with high density of Pgp-1 appear in later life, though this is more marked in the thymus of SJL, which also shows a higher relative density at an advanced age, than do BALB/c mice. Furthermore, the per cent of cells with high density of Pgp-1 is larger in thymuses of SJL than in BALB/c mice. The percentage of CD45+ thymocytes remains unchanged, as animals age. Thymocyte-membrane densities of CD-45 undergo age-related increases in both SJL and BALB/c. The per cent of cells with high density of CD-45 is similar in both strains. Individual variations in relative size of subpopulations in SJL mice of the same age are greater in old than in young mice; this increase in heterogeneity is manifested by increase in standard deviation. Corresponding significant changes have not been observed in BALB/c or C57BL/6 mice. Thus, we have detected an intrastrain variation which may reflect age-related effects of the impact of stochastic events.

Postnatal dietary fat influences mRNAS involved in myelination.

The synthesis and composition of myelin in the developing mouse central nervous system can be influenced by diet. Postnatal maternal fat intake altered nursing pup brain and liver fatty acid composition. Peak (day 21) proteolipid protein (PLP) and myelin basic protein (MBP) mRNA levels were reduced when pups were nursed by mothers fed a fat-free or 5% coconut oil diet. This effect was reversed by feeding a corn oil based diet. Oleic acid accounts for about 30% of myelin fatty acids. mRNA levels of stearoyl CoA desaturase (SCD), the rate-limiting step in oleic acid synthesis, increase in neonatal mouse brain. Postnatal maternal fat-free feeding reduced day 21 pup brain SCD and LDL receptor, but not apolipoprotein (Apo E) E mRNA levels. In contrast to brain, nursing pup hepatic SCD mRNA levels were induced, LDL receptor mRNA levels were unaffected and Apo E mRNA levels were reduced by postnatal maternal fat-free feeding. Myelin-specific mRNA levels are developmentally regulated and influenced by dietary fat. Neonatal brain SCD and LDL receptor mRNA levels are also altered by neonatal fat intake. The neonatal response to dietary fat is tissue-specific at the mRNA level.

The alpha 1, alpha 2, and alpha 3 subunits of GABAA receptors: comparison in seizure-prone and -resistant mice and during development.

Gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in brain, opens chloride channels through actions on GABAA receptors. We now report base and amino acid sequences of the alpha 1, alpha 2, and alpha 3 subunits from GABAA receptors of audiogenic seizure-prone (DBA/2J) and -resistant (C57BL/6J) inbred strains of mice. Inbreeding had fixed different alleles of the alpha 1 subunit in the two strains, giving five base differences in the cDNAs. None of these affected amino acid sequence, but one did create a NsiI restriction site potentially useful in mapping genomic DNA. No base or amino acid sequence differences between the strains were detected for the other two subunits. Northern blots revealed no apparent strain differences in message levels for these three subunits in whole brains of the mice at 3 weeks of age, the peak of seizure susceptibility in DBA/2J, but did reveal distinct regional and developmental patterns of expression among the subunits in mouse brain.

Idiotypic profile of natural autoantibodies in newborn and young adult BALB/c mice.

Idiotypic profiles of autoreactive monoclonal antibodies (MoAb) were evaluated by their reactivity with a panel of alkaline phosphatase (AP)-coupled detector MoAb derived from the same fusions. Attention was given to the question of whether differences exist between MoAb derived from spleen cells (SC) or thymocytes (TC) and whether ID profiles would change during post-natal development. In the newborn, natural autoantibodies and MoAb which did not react with any one of eight autoantigens displayed different ID profiles, autoreactive MoAb being characterized by the expression of a restricted pattern of ID. During post-natal development, changes of ID expression were only observed with autoreactive MoAb. Many ID which were detected on MoAb derived from 6-day-old mice were not detected on SC-derived MoAb from young adults, while a few ID were significantly over-represented. Furthermore, especially with TC-derived MoAb, a clear linkage between certain idiotypes and autoantigen specificities could be demonstrated. Thus, in contrast to non-autoreactive MoAb, natural autoantibodies in the young adult were characterized by expressing only a selected number of ID at high frequency. Furthermore, the B-cell environment apparently played a role, since there were marked differences between ID profiles of TC- versus SC-derived MoAb. The data are interpreted in the sense that expansion and maturation of naturally activated autoreactive B cells are controlled rather than being random processes.

Insulin-like growth factor I and II gene expression in Balb/C mouse liver during postnatal development.

We studied the developmental shift from IGF-II to IGF-I mRNA synthesis in Balb/C mouse liver. IGF-I mRNA levels were low at birth, increased with age to peak at weaning, and IGF-I gene expression again increased during puberty. IGF-II expression was high during early postnatal development, but became nondetectable at weaning. Thus in early life there is a reciprocal relationship of rising IGF-I and falling IGF-II mRNA. Northern analysis revealed four IGF-I mRNA transcripts ranging from 1.1-7.0 kb, with the 1.1 kb being the most prominent. Five IGF-II transcripts ranging from 1.7-4.0 kb were found, with the 4.0 kb being the most prominent. The pattern of Serum IGF-I and IGF-II values paralleled liver IGF-I and IGF-II mRNA levels.

Strain differences in mouse brain weight gain and spatial-location scores during postnatal development.

Brain weight and performance on a spatial-location test were measured from birth to weaning on mouse strains CD1, Balb/c, and their F1 cross. For CD1, onsets of both the last rapid-brain-growth stage and nonzero scores on the spatial-location test were around age 17 days; asymptotes of both were reached about age 23 days. Balb/c and F1 had at least 5- to 6-day delays of both onsets and asymptotes. For the strains studied, onset of nonzero performance on the spatial-location test may be a provisional indicator of onset of significant and rapid brain growth after age 15 days. The delayed F1 brain growth stage could indicate that control of brain growth about age 20 days resides in an inhibitory factor present in Balb/c but not in CD1.

Antigen-independent selection of T15 idiotype during B-cell ontogeny in mice.

Precursors of B cells capable of responding to a T-independent form of phosphorylcholine (PC) in splenic focus assays were detected in the spleens of neonatal mice as early as 4 days after birth. The earliest anti-PC B cells were T15-. T15+ foci-forming B cells were first detected 6 days after birth and expanded rapidly to constitute greater than 80% of the total PC-specific foci by day 10. Injection of heat-killed S. pneumoniae (R36A) into neonatal mice resulted in priming of the antibody response to PC, with an idiotype profile reflecting that of precursors of foci-forming B cells at the time of antigen administration. Priming of 2-day-old mice with 2 x 10(6) and 2 x 10(7) R36A induced a five- and ten-fold increase in the antibody response to phosphorylcholine 6 to 8 weeks later. However, only 10 to 15% of the serum antibodies expressed the normally dominant T15 idiotype. Doses below 2 x 10(5) R36A showed no detectable priming activity. PC-specific hybridomas derived from mice injected with 2 x 10(7) R36A 2 days after birth lacked the idiotypic and molecular characteristics typical of T15+ antibodies. Antibodies to phosphorylcholine, raised by immunization of 6-week-old mice are normally protective against pneumococcal infection. However, serum antibodies from mice treated with R36A 2 days after birth and responding to phosphorylcholine following challenge with R36A at 6 weeks of age failed to protect against deliberate infection with virulent S. pneumoniae. These observations imply that the antigen phosphorylcholine does not play a role in the selective expansion and dominant expression of the T15 idiotype.