Onset Of Lupus Nephritis Research Articles

Antagonist of monocyte chemoattractant protein 1 ameliorates the initiation and progression of lupus nephritis and renal vasculitis in MRL/lpr mice.

To examine whether chemokine antagonists inhibit the initiation and progression of lupus nephritis in MRL/lpr mice. NH(2)-terminal-truncated monocyte chemoattractant protein 1 (MCP-1)/CCL2 or thymus and activation-regulated chemokine (TARC)/CCL17 analogs were inserted into the pCXN2 expression vector and transfected into a nonmetastatic fibroblastoid cell line, MRL/N-1, established from an MRL/gld mouse. MCP-1 antagonist- or TARC antagonist-transfected MRL/N-1 cells were injected subcutaneously into MRL/lpr mice ages 7 weeks (before the onset of lupus nephritis) and 12 weeks (at the early stage of the disease). After 8 weeks, mice bearing the MCP-1 antagonist showed markedly diminished infiltration of macrophages and T cells, glomerular hypercellularity, glomerulosclerosis, crescent formation, and vasculitis compared with control mice. This seemed to be due to decreased production of interferon-gamma and interleukin-2 in the kidney. In contrast, there was no significant difference in renal damage between mice bearing TARC antagonist and control mice. We established a new system using MRL/N-1 cells that allows long-term observation of the effects of chemokine antagonists on lupus nephritis in MRL/lpr mice. We also showed that the MCP-1 antagonist ameliorated the initiation and progression of lupus nephritis and of renal vasculitis, which might provide a new approach to the treatment of the disease.

Intrathymic kidney cells delay the onset of lupus nephritis in MRL-lpr/lpr mice.

Introduction of antigens to maturing T cells in the neonatal thymus is an effective means of inducing tolerance; however, it is uncertain whether developing, pathogenic, autoreactive T cells can be selectively modulated during systemic autoimmunity. To address this issue, syngeneic cells, derived from the kidneys of pre-diseased, lupus-prone, MRL-lpr/lpr mice were administered intrathymically to either neonatal or young MRL-lpr/lpr mice. For comparison, littermates were injected with splenocytes, islet cells or saline. Kidney cells administered to neonatal mice resulted in attenuation of nephritis, despite elevated serum autoantibody levels, IgG deposits and lymphadenopathy. This effect was not observed with administration of either the renal cell preparations to older mice or islet cells to younger mice, although splenocytes provided some benefit in younger mice. The results indicate that a subset of autoreactive T cells, distinct from those that augment autoantibody production and lymphadenopathy, are necessary for the expression of severe nephritis in MRL-lpr/lpr mice, and they provide further support for a renal antigen-specific component to the phenotype,

Naturally processed chromatin peptides reveal a major autoepitope that primes pathogenic T and B cells of lupus.

Major autoepitopes for pathogenic Th cells of lupus were previously found in core histones of nucleosomes by testing overlapping synthetic peptides. To detect other dominant epitopes, we eluted peptides from MHC class II molecules of a murine lupus APC line that was fed with crude chromatin. The eluted peptides were purified by reverse-phase HPLC and tested for their ability to stimulate autoimmune Th clones, and then analyzed by mass spectrometry. Amino acid sequences of stimulatory fractions revealed three new autoepitopes. Two of the epitopes were homologous to brain transcription factor BRN-3, whereas the third sequence was homologous to histone H1'(22-42). H1'(22-42) stimulated autoimmune Th cells to augment the production of pathogenic antinuclear Abs, and was much more potent than other nucleosomal epitopes in accelerating glomerulonephritis in lupus-prone (SWR x NZB)F(1) (SNF(1)) mice. Remarkably, a marked expansion of Th1 cells recognizing the H1'(22-42) epitope occurred spontaneously in SNF(1) mice very early in life. A significant proportion of H1'(22-42)-specific T cell clones cross-reacted with one or more core histone epitopes, but not with epitopes in other lupus autoantigens. The H1'(22-42) epitope was also recognized by autoimmune B cells, and with the onset of lupus nephritis, serum autoantibodies to the H1'(22-42) epitope become increasingly cross-reactive with nuclear autoantigens. Convergence of T and B cell epitopes in H1'(22-42) and its ability to elicit a cross-reactive response make it a highly dominant epitope that could be targeted for therapy and for tracking autoimmune T and B cells.

Bone marrow transplantation: a new strategy for intractable diseases.

Bone marrow transplantation is becoming a powerful strategy for the treatment of hematologic disorders (leukemia, aplastic anemia, etc.), congenital immunodeficiencies, metabolic disorders and also autoimmune diseases. Using various animal models for autoimmune diseases, we have previously found that allogeneic (not autologous) bone marrow transplantation can be used to treat autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, immune thrombocytic purpura, insulin-dependent diabetes mellitus, chronic glomerulonephritis and certain types of non-insulin-dependent diabetes mellitus. In contrast, we have found that the transplantation of T-cell-depleted bone marrow cells or partially purified hemopoietic stem cells from autoimmune-prone mice to normal mice leads to the induction of autoimmune diseases in the recipients. These findings have recently been confirmed even in humans; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis and Crohn's disease were resolved after allogeneic bone marrow transplantation. However, there have recently been reports on the rapid recurrence or persistence of autoimmune diseases after autologous bone marrow transplantation. Conversely, the adoptive transfer of autoimmune diseases such as myasthenia gravis, insulin-dependent diabetes mellitus and Graves' disease by allogeneic bone marrow transplantation from donors to recipients has been reported. Owing to these findings, we have proposed that autoimmune diseases are "stem cell disorders." We have thus succeeded in treating autoimmune diseases in various autoimmune-prone mice, except MRL/lpr mice, by conventional bone marrow transplantation. The MRL/lpr mouse itself is radiosensitive (<8.5 Gy), while the abnormal hemopoietic stem cells of the MRL/lpr mouse are radioresistant (>8.5 Gy); conventional bone marrow transplantation (8.5 Gy plus bone marrow transplantation) has a transient effect on autoimmune diseases, which recur three months after the bone marrow transplantation. However, bone marrow transplantation plus bone grafts (to recruit donor stromal cells) completely prevents the recurrence of autoimmune diseases in MRL/lpr mice. Donor-derived stromal cells (including mesenchymal stem cells) thus seem to play a crucial role in successful allogeneic bone marrow transplantation, since there is a major histocompatibility complex restriction between hemopoietic stem cells and stromal cells. We have, however, found that the combination of bone marrow transplantation plus bone grafts has no effect on the treatment of autoimmune diseases in MRL/lpr mice, since MRL/lpr mice become more radiosensitive after the onset of lupus nephritis due to the development of uremic enterocolitis. To reduce the cytotoxic effect of radiation on the intestine, we carried out fractionated irradiation and devised a new strategy. We injected allogeneic whole bone marrow cells (including a small number [<3%] of T cells, hemopoietic stem cells and stromal cells) from donors directly into the intra-bone marrow of recipients so that donor-derived hemopoietic cells including stromal cells could effectively accumulate in the bone marrow. All the MRL/lpr mice survived more than one year (>60 weeks after birth) without the recurrence of autoimmune diseases, and immunological functions were completely restored even when the radiation dose was reduced to 5 Gy x 2. These findings suggest that intra-bone marrow injection-bone marrow transplantation can be used to treat intractable autoimmune diseases under reduced radiation doses without using any immunosuppressants.Intra-bone marrow injection-bone marrow transplantation seems to be the best strategy for allogeneic bone marrow transplantation: 1) no graft-versus-host disease develops even if T cells are not depleted from the bone marrow; 2) no graft failure occurs even if the dose of radiation as the conditioning for bone marrow transplantation is reduced to 5 Gy x 2; 3) hemopoietic recovery is rapid; and 4) T-cell functions are completely restored even in donor-recipient combinations across the major histocompatibility complex barriers. Using cynomolgus monkeys, we have recently established a new method (the "perfusion method") for collecting bone marrow cells from the long bones (femur, humerus, etc.) without peripheral blood contamination. This method has various advantages: 1) no graft-versus-host disease develops even in cynomolgus monkeys, since the percentage of T cells in the bone marrow cells collected is less than 3%; 2) a large number of bone marrow cells can be collected quickly and safely; and 3) the bone marrow cells collected contain stromal cells including mesenchymal stem cells. We therefore believe that this method (intra-bone marrow injection-bone marrow transplantation in conjunction with the perfusion method) will become a powerful new strategy for not only allogeneic bone marrow transplantation but also organ transplantation in conjunction with bone marrow transplantation. Furthermore, this method could become a valuable strategy in regeneration therapy for injured organs and tissues (myocardial infarction, cerebral infarction, Alzheimer's disease, etc.), since it can efficiently reconstitute the recipient with both donor-derived hemopoietic stem cells and mesenchymal stem cells.

Treatment of autoimmune diseases by hematopoietic stem cell transplantation.

Remarkable advances have been made in bone marrow transplantation (BMT), which now has become a powerful strategy for the treatment of leukemia, aplastic anemia, congenital immunodeficiency disorders, and autoimmune diseases. Using various animal models, allogeneic BMT has been found to be useful in the treatment of autoimmune diseases. In MRL/lpr mice, which are radiosensitive (<8.5 Gy) and are an animal model for autoimmune disorders, conventional BMT resulted in only transient effects; the manifestations of the autoimmune diseases recurred 3 months after BMT. However, the combination of BMT plus bone grafts (to recruit donor stromal cells) was capable of preventing the recurrence of autoimmune diseases in MRL/lpr mice. This strategy was found to be ineffective in the treatment of MRL/lpr mice that had developed autoimmune diseases, because these mice were more sensitive to the effects of radiation after the onset of lupus nephritis due to uremic enterocolitis. We have recently discovered a safer strategy for treatment of autoimmune diseases, which includes fractionated irradiation (5.5 Gy x 2) (day -1) followed by portal venous injection (day 0) plus intravenous injection (day 5) of donor unfractionated bone marrow cells. We successfully treated autoimmune diseases in MRL/lpr mice using this strategy; 100% of MRL/lpr mice treated in this fashion survive >1 year after treatment. We identified the mechanisms underlying the components of this approach and have found that stromal cells play a crucial role in successful BMT. In this review, the conditions essential for successful allogeneic BMT are discussed.

Treatment of autoimmune disorders by hemopoietic cell transplantation

Abstract We have previously found that allogeneic BMT can be used to treat autoimmune diseases using various animal models for autoimmune diseases (Proc. Natl. Acad. Sci. USA 82:2483, 1985, etc.). However, in MRL/lpr mice that are radiosensitive (less than 8.5Gy), conventional BMT was found to have transient effects; the autoimmune diseases recur 3 months after BMT. However, we have found that the combination of BMT plus bone grafts (to recruit donor stromal cells) completely prevents the recurrence of autoimmune diseases in these mice (J. Immunol. 152: 3119, 1994). In addition, we have found that this strategy has no effect on the treatment of autoimmune diseases in MRL/lpr mice, since these mice become more radiosensitive after the onset of lupus nephritis, due to uremic enterocolitis. We have very recently discovered a new safe strategy for the treatment of autoimmune diseases which includes fractionated irradiation (5.5 Gy × 2) (day−1) followed by portal venous (P.V.) injection (day 0) plus intravenous (I.V.) injection (day 5) of donor bone marrow cells. We have succeeded in treating autoimmune diseases in MRL/lpr mice by this strategy; 100% of thus-treated MRL/lpr mice survive more than one year after the treatment (Blood 95: 1862, 2000). We have also found that this strategy is applicable to organ allografts in mice; all the skin and pancreas allografts survive more than one year after transplantation without using immunosuppressants. We clarify the mechanisms underlying the advantages of this strategy, and also show using cynomolgus monkeys that this strategy will become applicable to humans.

Lessons from the NZM2410 Model and Related Strains

SLE susceptibility requires the interplay of an unknown number of genes and equally unidentified triggering events. The past few years have seen significant advances in our understanding of SLE susceptibility through the genetic analysis of murine models. The NZM2410 strain, which is derived from the NZB/WF1 model has played a significant role in these advances. The main advantages presented by this strain over other models are the genetic homozygozity at all loci and an highly penetrant early onset lupus nephritis in both males and females, indicating that the strongest BWFj susceptibility loci were retained in NZM2410. After identification of NZM2410 susceptibility loci via linkage analyses, congenic strains have been derived in order to convert a polygenic system into a series of monogenic traits. These congenic strains have been analyzed in an integrated process which has provided simultaneously 1) novel functional characterization of the Sle susceptibility loci, 2) high resolution genetic maps that will lead to the identification of the corresponding susceptibility genes by either candidate locus or positional cloning, and 3) insights into the mechanisms by which these loci interact to produce systemic autoimmunity with fatal end-organ damage.

Reciprocal T-B determinant spreading develops spontaneously in murine lupus: implications for pathogenesis.

Recent work from several laboratories has shown that, in contrast to the widely held notion that one autoimmune disease is caused by one or a few related autoantigenic determinants, autoimmunity is fundamentally a continuously evolving process. The autoimmune responses shift, drift and diversify with time not only to other determinants in the original antigen but also to other antigens. We have described a form of determinant spreading--reciprocal T-B determinant spreading--where the induction of first T cells by peptides from an autoantibody molecule could lead to help provided to a variety of B cells displaying a cross-reactive version of the original determinant. The response spreads in this way by reciprocal T-B stimulation until large cohorts of T and B cells have expanded. Such spontaneous expansion must be important in clinical disease, since tolerance induction to a limited set of T-cell determinant peptides derived from an anti-DNA antibody VH region delayed the appearance of IgG anti-dsDNA antibodies and onset of lupus nephritis in the NZB/NZW F1 mouse model of systemic lupus erythematosus. Understanding the diversification patterns in autoimmune responses has enormous implications in developing peptide-targeted therapies.

INTRARENAL T CELLS IN LUPUS NEPHRITIS ANALYZED BY CDR3 SIZE SPECTRATYPING♦ 1842

Increasing evidence supports a pivotal role of T cells in the pathogenesis of a number of autoimmune processes. Lupus nephritis (LN) is characterized by variable degrees of glomerular and interstitial inflammation comprised primarily of lymphocytes and plasma cells. We have used the process of CDR3 spectratyping to analyze the TCR Vβ repertoire complexity based on CDR3 length heterogeneity of intrarenal T cells in newly diagnosed LN as compared to corresponding peripheral blood. Following PCR amplication of cDNA with each of 25 Vβ oligonucleotide 5′ primers paired with a single fluorescently labeled 3′ Cβ primer, CDR3 spectratypes were generated through the analysis of these products on denaturing acrylamide gels. For evaluation of spectratypes, histograms were generated from data collected using an automated DNA sequencer and GeneScan Analysis software. Analysis of 4 patients with new onset LN demonstrated that intrarenal T lymphocytes contain oligoclonal expansion as compared to circulating T lymphocytes. Thus, the method of CDR3 spectratyping provides a more indepth analysis of the TCR repertoire and is a powerful tool to search for antigen-driven clonal expansions.Supported by National Kidney Foundation

Roles of IL-4 and IL-12 in the development of lupus in NZB/W F1 mice.

Development of either Th1 or Th2 cell subsets has profound immunologic consequences, either pathogenic or protective, in several autoimmune diseases. However, it remains unclear which subset of Th cells plays a more critical role in lupus. In this study, we examined IL-4 and IL-12, which play decisive roles in the development of Th2 and Th1, respectively, in the IgG autoantibody production and development of lupus nephritis in NZB/W (B/W) F1 mice. Transfer of either IL-4- or IL-12-stimulated splenocytes from 5-mo-old B/W F1 mice into B/W F1 mice of the same age enhanced the production of IgG anti-dsDNA Ab. Consistently, administration of mAb against either IL-4 or IL-12 before the onset of lupus could inhibit the production of IgG anti-dsDNA Ab. However, only anti-IL-4 mAb was effective in preventing the onset of lupus nephritis. This discrepancy appeared to be explained by the differential effect on the production of IgG3-type autoantibody and TNF production. Interestingly, when combined, anti-IL-12 mAb abrogated the beneficial effect of anti-IL-4 mAb. These results indicate that both Th2 and Th1 contribute to the IgG autoantibody production, and IL-4 and IL-12 play key roles in the complexity of cytokine regulation in the pathogenesis of autoimmunity in lupus, but the former is more critical.

Clinical Renal Involvement in Afro-Caribbean Lupus Patients

This study reports the severity of clinical lupus nephritis (LN), defined as the sustained presence of proteinuria and/or cellular casts, in a group of 68 newly diagnosed patients with systemic lupus erythematosus (SLE) in Curaçao. Fifty-four patients (78%) developed clinical signs of LN, of whom 31 (54%) had clinical LN at the time of SLE diagnosis. The probability of developing clinical LN reached 80% in the first 3 years after SLE diagnosis and hardly increased later. No clinical or serological differences existed at the time of SLE diagnosis or at onset of LN between patients with early-or late-onset LN. Survival in patients without LN was 100% at 5 years, while for patients with clinical LN these rates at 1 and 5 years were 91% and 59%, respectively (P = 0.0001); male LN patients had a worse prognosis than females (P = 0.012), while time of LN onset did not influence survival. Six patients (11%) developed end-stage renal failure; all were female, five had early LN and one had late-onset LN (P = 0.17). Renal survival was 97% and 80% at 1 and 5 years, with decreased rates for patients with nephrotic-range proteinuria (P = 0.02). Hypertension was present in 13% of LN patients, but had no influence on patient or renal survival. Thus, clinical LN was a frequent complication, which carried a poor prognosis in these Afro-Caribbean lupus patients.

A case of progressive encephalomyelitis with rigidity and positive antiglutamic acid decarboxylase antibodies [corrected

A 50 year old woman developed progressive encephalomyelitis with rigidity over a three year period. Her CSF contained oligoclonal bands and both her serum and CSF contained antibodies directed against GABA-ergic synapses (antiglutamic acid decarboxylase [corrected] antibodies). These antibodies have recently been described in cases of stiff man syndrome. Both disorders may be part of a clinical spectrum that has an underlying autoimmune basis.

ＹＭ‐１３６５０の実験腎炎に対する作用

We investigated the effects of YM-13650, 2-(m-carboxyacetoxyphenyl) imidazo [2, 1-b] benzothiazole, on BSA-immune complex nephritis in rats and lupus nephritis in NZB/W F1 mice. In preventative experiments on immune complex nephritis in rats, YM-13650 (10 approximately 100 mg/kg, p.o.) dose-dependently inhibited the increase in urinary protein, serum cholesterol, and urea nitrogen. Histopathological observation showed striking hypercellularity and mesangial widening in nephritic control; however, glomerular injury was reduced in YM-13650-treated animals. In therapeutic study, control rats maintained high levels of urinary protein, serum cholesterol and urea nitrogen throughout the experimental period. These variables were lower in YM-13650-treated rats. In preventative experiments in lupus mice treated from 8 weeks of age, YM-13650 in comparison with the control group showed a lesser degree of proteinuria throughout the experimental period. It also significantly prolonged or tended to prolong the life span of NZB/W F1 mice compared with the control. In therapeutic experiments conducted after the onset of lupus nephritis in mice, the drug also inhibited an increase in urinary protein and tended to prolong the life span. These results show that YM-13650 has preventative and therapeutic effects on experimental nephritis in rats and mice, and it may prove valuable as an anti-nephritic drug.