Genetic control of the immune response to ferritin in F1 hybrid mice.

The immune response to the antigen horse spleen ferritin, has been investigated in ten inbred parental strains and seven different F1 hybrid strains of mice, using an antigen excess technique. The degree of dominance in an F1 hybrid system can be estimated by using the Fisher dominance index. The responses in F1 hybrid animals, obtained from crosses of high and low responder parents, varied from dominant to recessive but the overall mean dominance index for the ferritin immunogenetic system was found to be -0.0467 +/- 0.0083 (mean +/- s.e.), a value close to zero, which suggests a codominant mode of inheritance of IR-genes to ferritin and this is consistent with most published data in other F1 IR-gene systems.     

Mediators of joint swelling and damage in rheumatoid arthritis and pristane induced arthritis.

Joint swelling and tenderness in rheumatoid arthritis (RA) probably result from IgG aggregates activating complement with the consequent attraction of polymorphonuclear leucocytes (PMNs) and the liberation of their granule enzymes such as kininogenases. By contrast IL-1 and TNF are the major stimulants of cartilage and bone loss although other agents contribute. The fundamental drive for the production of these mediators is unknown but a role for heat shock proteins is suggested from work on pristane induced arthritis.     

Dietary frying oil influences immune regulation in autoimmune-prone NZBxNZW F1 mice

To further elucidate the role of dietary frying oil in the pathogenesis of autoimmune diseases, two groups of NZB/W F1 mice were fed with diets containing 20% fresh oil and frying oil, respectively. All these mice were followed up serum anit-DNA antibody levels, proteinuria and life span regularly. Our data suggested: 1) higher IgG anti-ss, dDNA antibody levels were noted in mice fed with fresh oil compared to those of the frying oil group; 2) lipopolysaccharide (LPS)-stimulated spleen cells of mice fed with frying oil produced higher IL-10 compared to that of fresh oil group; 3) IL-6, TNF-alpha and PGE2 produced by macrophages of dietary frying oil group were higher, although not statistically significant, than those of fresh oil group. Different degree of deterioration of dietary oil has been found to affect immune response in autoimmune mice.     

Life-long tolerance and chimerism in parental mice induced with F 1 hybrid cells

We have induced specific immunological tolerance towards (C57BL × CBA-T6T6) F₁ skin grafts in newborn recipients of the parental CBA strain by one injection of 20 × 10⁶ or 40 × 10⁶ F₁ spleen cells. Approximately 80 % of the CBA recipients were highly tolerant (the grafts survived for more than 50 days). Many of the highly tolerant mice retained fully preserved grafts until natural death. The highly tolerant CBA mice had dividing (C57BL × CBA-T6T6) F₁ cells in their spleen (12–13 %) and lymph nodes (12–15 %). The proportion of these cells decreased with time, but we found a few of them even at 567 days of life (0.7 and 4.2 %, respectively). We have concluded that specific immunological tolerance to tissue grafts induced at birth can last throughout the entire life-span of a mouse. The tolerance persists in spite of a very low proportion of dividing donor cells within the lymphoid tissue of tolerant hosts.     

Genetic control of inflammatory arthritis and glomerulonephritis in congenic lpr mice and their F1 hybrids.

MRL-lpr/lpr mice spontaneously develop a complex immunological disease characterized by glomerulonephritis, inflammatory erosive arthritis and the production of rheumatoid factors (RF) and anti-DNA antibodies. We have previously reported that, of congenic lpr strains, only MRL-lpr/lpr mice develop synovial pathology suggesting that both the lpr gene and another gene(s) in the MRL background are necessary for the development of arthritis. To define further the genetics of arthritis and its relationship to glomerulonephritis and autoantibody production, we studied disease expression in MRL-lpr/lpr and C57BL/6-lpr/lpr mice and their offspring (BM-lpr/lpr and MB-lpr/lpr). At 6 months of age these mice were killed and bled, and their kidneys and knee joints were removed for pathological studies. Fourteen of 28 MB-lpr/lpr mice displayed synovial hypertrophy, while eight of 28 had significant synovial inflammation. BM-lpr/lpr mice showed similar changes: nine of 22 and eight of 22 exhibited synovial hypertrophy and inflammation respectively. Joints from MRL-lpr/lpr mice revealed 13 of 17 with synovial hypertrophy and 12 of 17 with inflammation, while none of 14 B6-lpr/lpr mice had synovial changes. Renal pathology was minimal in the F1 mice with only mild hypercellularity in seven of 21 MB-lpr/lpr and five of 22 BM-lpr/lpr mice. All MRL-lpr/lpr mice, in contrast, had marked glomerular changes with 12 of 17 exhibiting glomerular crescents. Only one F1 mouse had both arthritis and renal abnormalities. IgM RF levels were elevated in all four experimental groups, but did not correlate with the presence or severity of arthritis. IgG RF levels were elevated in the MB-lpr/lpr and MRL-lpr/lpr mice, but did not correlate with the degree of arthritis. These results indicate that renal disease and arthritis develop independently in lpr mice, possibly on a genetic basis, and that the presence and titer of autoantibodies do not correlate with tissue injury.     

Proteoglycan-induced arthritis: immune regulation, cellular mechanisms, and genetics

Rheumatoid arthritis is probably the least understood systemic autoimmune disease, and it affects approximately 1% of the human population. Several lines of evidence indicate that the effector mechanism, which initially attacks small joints, is T-cell driven. As a result, an aggressive synovial pannus develops, which destroys articular cartilage and bone, leading to massive ankylosis and deformities of peripheral joints. The disease has a progressive character, with the involvement of more and more joints. Although the target organ is the synovial joint, there is no clear evidence that any macromolecule of cartilaginous tissues, bone, or synovium, could be a preferential autoantigen. There are numerous rodent models that simulate some or many of the clinical, immunological, or histopathological features of the disease. Recently, it has become a strong working hypothesis that MHC and non-MHC genetic components share loci that are common in various autoimmune diseases, and in corresponding animal models. The most relevant animal models of rheumatoid arthritis appear to be those induced by cartilage matrix components such as type II collagen or proteoglycan aggrecan. This review summarizes our current knowledge of cartilage proteoglycan (aggrecan)-induced arthritis in mice.     

Macrophage clearance function and immune complex disease in New Zealand Black/White F1 hybrid mice.

Macrophage clearance function in NZB, NZW, NZB/W, Ajax and B10D2 new mice was assessed by measurement of the rate of clearance (KPVP) of intravenously-injected 125I-labelled polyvinylpyrrolidone (PVP). There were significant strain and age-related variations in KPVP. In particular there was a marked fall in KPVP in NZB/W mice with increasing age. This fall was most apparent in female NZB/W and preceded the age at which renal disease usually develops in these animals. We suggest that ineffective macrophage function and production of low affinity antibody contribute to the early development of immune complex glomerulonephritis in these mice.     

Age- and sex-dependent thymic abnormalities in NZB × SJL F1 hybrid mice

The cellular organization of the thymus was investigated in 3- and 12-month-old NZB × SJL F₁ hybrid (NS) mice. Age-dependent alterations were demonstrated which differed strikingly according to the sex of the animals. In female mice, marked abnormalities of the thymus developed during ageing. They consisted of a more or less pronounced hypertrophy accompanied by histological changes and modifications in the nature of the lymphocyte populations. Three types of qualitative changes were found at 12 months of age: (1) depletion of cortical thymocytes as evidenced by histology, by the evaluation of peanut-agglutinin (PNA) binding and by cell electrophoresis; (2) hyperplasia of the medullary lymphoid tissue, probably reflecting the expansion of a population of mature T lymphocytes. This was further suggested by a rise (up to 60%) in the frequency of lymphocytes lacking both PNA receptor and B cell markers, by an increased proportion (57%) of high electrophoretic mobility (EPM) lymphocytes and by an augmentation of in vitro reactivities to phytohaemagglutinin (PHA) and, although to a lesser extent, to concanavalin A (Con A). (3) The appearance of significant numbers of B lymphocytes (up to 20%) as assessed by surface immunoglobulin (sIg) and complement receptor (CR) detection which was accompanied by a vigorous responsiveness of thymus cells to lipopolysaccharide (LPS). None of these abnormalities was seen in the male mice. Instead, the thymus of NS males displayed a nearly normal age-related involution without major change in the proportions of its lymphocyte subpopulations. NS mice thus provide an interesting model of thymic disease influenced by sex-linked factors.     

The immune response in NZB × NZW F1 hybrid mice

A study was made of the immune response of NZB × NZW F₁ hybrid (BW) mice against `H' antigen of Salmonella, sheep red blood cells, bovine serum albumin and allogeneic tumour cells. The responses of old BW mice having autoimmune glomerulonephritis were compared with those of young healthy mice. It was found that the mice develop immune depression concurrently with the autoimmune process. In old BW mice, the immune response is of the same type as the response observed in neonatally thymectomized mice. Neither age alone, nor functional deficiency of the thymus could account for this immune depression. It is suggested that the ability to respond to foreign antigens was probably depressed by the competition provided by autoantigens.     

Reduction of graft-versus-host disease in neonatal F1 hybrid mice.

Pre-immunization of BALB/c (H-2d) mothers with C57BL/10 (H-2b) or CBA/H (H-2k) spleen cells partially protected the F1 hybrid offspring of (BALB/c x C57BL/10) or (BALB/c x CBA/H) matings from graft-versus-host-disease (GVHD) induced by neonatal intraperitoneal inoculation with spleen cells of the paternal strain. The effects achieved were manifest as a reduction in mortality. Experiments to establish whether the phenomenon was antibody mediated were performed by passive pre-immunization of BALB/c mothers with alloantisera obtained from BALB/c previously immunized with C57BL/10 spleen cells. Alloantisera produced an equivalent reduction in GVHD mortality. Some of the F1 mice that survived challenge with paternal strain spleen cells were proven to be haemopoietic chimaeras using immunofluorescence with anti-MHC monoclonal antibodies and polymorphism of the enzyme glucose-phosphate-isomerase present in the strains used. The possible mechanisms of protection from GVHD in our mouse model are discussed.     

Cellular and humoral reactivity pattern to the mycobacterial heat shock protein hsp65 in pristane induced arthritis susceptible and hsp65 protected DBA/1 mice.

We have analysed the cellular and humoral immunity to the mycobacterial 65 kD heat shock protein (hsp65) in groups of DBA/1 mice with arthritis induced by intraperitoneal injection of the mineral oil pristane. Here we confirm that DBA/1 mice are highly susceptible to pristane induced arthritis (PIA) and demonstrate that the incidence of arthritis can be modulated by either pretreatment with low dose irradiation or by preimmunisation with recombinant hsp65. Global cellular responses to antigens such as BSA or type II collagen were not enhanced or impaired within groups of arthritic (A) or non-arthritic (NA) mice. However, the cellular response to hsp65 in arthritic animals preimmunised with the 65 kD antigen was significantly elevated in comparison to hsp65 preimmunised mice that were resistant to the induction of disease. On the contrary, the level of hsp65 specific antibodies was much high in NA animals than in PIA mice. CBA/Igb mice are partially susceptible to the induction of PIA. We have previously reported that arthritic CBA/Igb mice have both elevated cellular and humoral reactivity to hsp65. Although a central pivotal role for hsp65 has been postulated in autoimmune diseases these results indicate that there is no simple relationship between the pathogenesis of PIA and immune responses to hsp65.     

Studies on type II collagen induced arthritis in mice

A consistent and reproducible polyarthritis was induced in mice by immunizing them with type II collagen in Complete Freunds adjuvant (CFA) and Bacillus Calmette-Guerin (BCG) vaccine. Several inbred strains of mice were investigated for the ability to develop collagen induced arthritis (CIA). DBA/1 mice (H-2q) produced the highest incidence and the most severe arthritis of all the strains examined. Viable BCG vaccine was essential for the induction of a reproducible disease in this strain. The effects of some anti-inflammatory and anti-rheumatic compounds were examined on the developing and established lesions of CIA. These effects were determined by assessing the paw inflammation using a subjective scoring system and measuring foot weight. Furthermore, levels of serum amyloid P component (SAP) were also determined.Benoxaprofen, cyclophosphamide, indomethacin and prednisolone inhibited the paw inflammation in the developing disease whilst the anti-rheumatic compounds auranofin and D-penicillamine exacerbated the paw inflammation. Cyclophosphamide and prednisolone inhibited the established lesions but only prednisolone prevented the development of further lesions in the established disease. The SAP levels in the prednisolone treated group were also reduced. Auranofin treatment exacerbated the inflammation of both the established and the developing lesions in the same animal. D-penicillamine was inactive in the established disease.     

丙型肝炎病毒核心基因疫苗诱导小鼠细胞及体液免疫应答

为了探索丙型肝炎病毒核心区基因疫苗对小白鼠细胞及体液免疫功能的影响。本文将ＨＣＶ核心区基因定向克隆于质粒ｐｃＤＮＡ３巨细胞病毒启动子下游,构建了能在真核细胞内表达的重组基因疫苗ｐｃＤＮＡ３－ＨＣＶ,将此疫苗通过多点肌肉注射免疫ＢＡＬＢ／Ｃ小鼠。结果发现接种了该疫苗的小鼠出现抗－ＨＣＶＩｇＧ阳性,其抗体水平明显高于对照组,同时Ｔｈ１免疫应答相关的细胞因子ＩＬ－２及γ干扰素水平也明显高于对照组,其结果     

A new model of AA-amyloidosis induced by oral pristane in BALB/c mice.

Fifteen male BALB/c mice were given six intermittent oral doses of O.I. ml pristane (2, 6, 10, 14 tetramethylpentadecane) within a period of 9 weeks. Fifteen mice receiving tap water using the same schedule formed the control group. Amyloidosis was first detected in the spleen of a mouse which had died 33 weeks after the first dose and 24 weeks after the last. All six mice which were subsequently autopsied 34-51 weeks after the first dose also showed amyloidosis involving liver and spleen. The most extensive tissue deposits were seen at 37-38 weeks whereas the older mice showed predominantly chronic renal lesions with papillary necrosis, scars and cystic change. Electron microscopy confirmed the identity of the amyloid fibrils and the presence of globular stellate amyloid ''bodies'' in liver and spleen. The amyloid deposits were shown to be made up of AA (amyloid associated) protein using an indirect immunoperoxidase method and a monoclonal rat anti-murine AA protein antibody. We did not find any plasmacytomas or increased numbers of plasma cells in the bone marrow. None of the control mice developed amyloidosis. This new experimental model promises to provide a means of studying several aspects of secondary amyloidosis which may be relevant to the clinical situation.     

Genetic influences on the immune repertoire following tuberculous infection in mice

Differences in the antibody repertoire between B10 and BALB background strains have been found following intraperitoneal infection of mice with Mycobacterium tuberculosis. Western blot analysis showed that B10 sera reacted with only a few antigenic bands, whereas BALB sera reacted with multiple bands, irrespective of the H-2 (b or k) haplotype. The oligo-banded pattern was a feature of live infection, since immunization with the mycobacterial extract in incomplete Freunds' adjuvant (IFA) produced a multi-banded response in both intact and in previously infected B10 mice. The multibanded BALB phenotype was dominantly expressed in (BALB x B10) F1 hybrids. The extent to which antibody levels to individual antigens varied was influenced by the combined effects of background (non-H-2) and H-2 genes: anti-65,000 molecular weight (MW) and anti-71,000 MW IgG levels were high in BALB but absent or low in B10 mice, irrespective of H-2 haplotypes, anti-19,000 MW levels in B10 mice were strongly H-2 controlled, whilst anti-38,000 MW levels were high in all tested strains. Immunoglobulin G1 (IgG1) subclass antibody and splenic interleukin-4 (IL-4) mRNA levels were distinctly lower in B10 than in BALB mice, but in vitro T-cell proliferative responses to mycobacterial antigens did not differ between the tested strains. It is proposed that the limited extracellular release of mycobacterial antigens required to stimulate B cells and/or differential activation of T-cell subsets may explain the narrow antibody repertoire in B10 strains of mice. This may have relevance to the outcome of infection, as bacterial counts were higher in the BALB strains at 10 weeks post-infection.     

Primary cardiac hemangiosarcomas induced by 1,3-butadiene in B6C3F1 hybrid mice

Proliferative vascular lesions of the heart were found in mice exposed chronically to 1,3-butadiene by inhalation with an overall incidence of 30% in males and 43% in females. Based on histological criteria, the lesions were subclassified as endothelial hyperplasia with an incidence of 7% in males and 13% in females and hemangiosarcoma with an incidence of 23% and 30%, respectively. A dose-relationship for both lesions was observed in females, but not in males. The absence of a dose response in males was most likely due to the lower survival rate for high-dose animals (14%) when compared to the lower-dose animals (22%). Endothelial hyperplasia was characterized by widened vascular spaces lined by a single layer of plump endothelial cells. When cellular pleomorphism and piling up of endothelial nuclei were observed, the lesion was diagnosed as hemangiosarcoma. Ultrastructural examination of hemangiosarcomas revealed lumen formation, intercellular junctions and cytoplasmic filaments. Pinocytotic vesicles which are 1 of the characteristics of endothelial cells could not be identified with certainty. Weibel-Palade bodies were not detected in the neoplastic endothelium. Metastatic lesions were observed in liver, lung and kidney. To date, 1,3-butadiene is the only carcinogen reported that induces proliferative vascular lesions in the heart of mice.     

Sex differences in the regulation of experimentally induced autoantibodies in (NZB x NZW)F1 mice.

The ability to induce autoantibodies to erythrocytes in male and female (NZB x NZW)F1 mice was examined. Female (NZB x NZW)F1 mice were shown to produce significantly more autoantibody than the male (NZB x NZW)F1 mice. The regulation of this experimentaly induced autoantibody was studied by examining the ability of male and female (NZB x NZW)F1 mice to generate antigen-specific suppressor cells. A sex difference was found in the ability to generate these suppressor cells. Male mice generated antigen-specific suppressor cells in response to rat RBC which were capable to suppressing the experimental induction of red cell autoantibodies whereas female mice were unable to generate those antigen-specific suppressor cells.