Explore the vast range of titles available.

Real-time reverse transcription PCR (RT-qPCR) has emerged as an accurate and widely used technique for expression profiling of selected genes. However, obtaining reliable measurements depends on the selection of appropriate reference genes for gene expression normalization. The aim of this work was to assess the expression stability of 15 candidate genes to determine which set of reference genes is best suited for transcript normalization in citrus in different tissues and organs and leaves challenged with five pathogens (Alternaria alternata, Phytophthora parasitica, Xylella fastidiosa and Candidatus Liberibacter asiaticus). We tested traditional genes used for transcript normalization in citrus and orthologs of Arabidopsis thaliana genes described as superior reference genes based on transcriptome data. geNorm and NormFinder algorithms were used to find the best reference genes to normalize all samples and conditions tested. Additionally, each biotic stress was individually analyzed by geNorm. In general, FBOX (encoding a member of the F-box family) and GAPC2 (GAPDH) was the most stable candidate gene set assessed under the different conditions and subsets tested, while CYP (cyclophilin), TUB (tubulin) and CtP (cathepsin) were the least stably expressed genes found. Validation of the best suitable reference genes for normalizing the expression level of the WRKY70 transcription factor in leaves infected with Candidatus Liberibacter asiaticus showed that arbitrary use of reference genes without previous testing could lead to misinterpretation of data. Our results revealed FBOX, SAND (a SAND family protein), GAPC2 and UPL7 (ubiquitin protein ligase 7) to be superior reference genes, and we recommend their use in studies of gene expression in citrus species and relatives. This work constitutes the first systematic analysis for the selection of superior reference genes for transcript normalization in different citrus organs and under biotic stress.

Autosomal recessive juvenile parkinsonism (AR–JP), one of the most common familial forms of Parkinson disease, is characterized by selective dopaminergic neural cell death and the absence of the Lewy body, a cytoplasmic inclusion body consisting of aggregates of abnormally accumulated proteins 1 . We previously cloned PARK2 , mutations of which cause AR–JP (ref. 2 ), but the function of the gene product, parkin, remains unknown. We report here that parkin is involved in protein degradation as a ubiquitin-protein ligase collaborating with the ubiquitin-conjugating enzyme UbcH7, and that mutant parkins from AR–JP patients show loss of the ubiquitin-protein ligase activity. Our findings indicate that accumulation of proteins that have yet to be identified causes a selective neural cell death without formation of Lewy bodies. Our findings should enhance the exploration of the molecular mechanisms of neurodegeneration in Parkinson disease as well as in other neurodegenerative diseases that are characterized by involvement of abnormal protein ubiquitination, including Alzheimer disease, other tauopathies, CAG triplet repeat disorders and amyotrophic lateral sclerosis 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 .

Background There is growing evidence that neuroinflammation may contribute to schizophrenia neuropathology. Elevated pro-inflammatory cytokines are evident in the midbrain from schizophrenia subjects, findings that are driven by a subgroup of patients, characterised as a “high inflammation” biotype. Cytokines trigger the release of antibodies, of which immunoglobulin G (IgG) is the most common. The level and function of IgG is regulated by its transporter (FcGRT) and by pro-inflammatory IgG receptors (including FcGR3A) in balance with the anti-inflammatory IgG receptor FcGR2B. Testing whether abnormalities in IgG activity contribute to the neuroinflammatory abnormalities schizophrenia patients, particularly those with elevated cytokines, may help identify novel treatment targets. Methods Post-mortem midbrain tissue from healthy controls and schizophrenia cases ( n  = 58 total) was used to determine the localisation and abundance of IgG and IgG transporters and receptors in the midbrain of healthy controls and schizophrenia patients. Protein levels of IgG and FcGRT were quantified using western blot, and gene transcript levels of FcGRT, FcGR3A and FcGR2B were assessed using qPCR. The distribution of IgG in the midbrain was assessed using immunohistochemistry and immunofluorescence. Results were compared between diagnostic (schizophrenia vs control) and inflammatory (high vs low inflammation) groups. Results We found that IgG and FcGRT protein abundance (relative to β-actin) was unchanged in people with schizophrenia compared with controls irrespective of inflammatory subtype. In contrast, FcGRT and FcGR3A mRNA levels were elevated in the midbrain from “high inflammation” schizophrenia cases (FcGRT; p  = 0.02, FcGR3A; p  < 0.0001) in comparison to low-inflammation patients and healthy controls, while FcGR2B mRNA levels were unchanged. IgG immunoreactivity was evident in the midbrain, and approximately 24% of all individuals (control subjects and schizophrenia cases) showed diffusion of IgG from blood vessels into the brain. However, the intensity and distribution of IgG was comparable across schizophrenia cases and control subjects. Conclusion These findings suggest that an increase in the pro-inflammatory Fcγ receptor FcGR3A, rather than an overall increase in IgG levels, contribute to midbrain neuroinflammation in schizophrenia patients. However, more precise information about IgG-Fcγ receptor interactions is needed to determine their potential role in schizophrenia neuropathology.

Background:The efficacy of mepolizumab (MPZ), an anti-IL-5 antibody for eosinophilic granulomatosis with polyangiitis (EGPA), has been reported. MPZ was approved for relapsing or refractory EGPA in 2018. We have reported the real-world effectiveness and safety of MPZ for relapsing and refractory EGPA in the maintenance phase (n=16) and in the remission induction phase (n=7). However, an evaluation with an extended observation period with a larger sample size is expected.Objectives:We evaluated the effectiveness and safety of MPZ (1) in the maintenance phase for relapsing and refractory EGPA (study 1) and (2) in the remission induction phase for severe EGPA (study 2) in 48 patients with EGPA in real clinical practice.Methods:This study consists of study 1(remission maintenance phase) and 2 (remission induction phase). Study 1: The 2-year effectiveness and safety in the maintenance phase of relapsing and refractory EGPA (N=33) were evaluated. Clinical data of 2 years before (Pre-MPZ group: month -24 to 0) and after (Post-MPZ group: month 0 to 24) MPZ introduction were compared. The primary endpoint was the remission-achieving rate in the Post-MPZ group (remission: BVAS 0 and GC[PSL] ≤ 4 mg/day). Secondary endpoints were GC-free remission rate, MPZ retention rate, BVAS, eosinophil counts (/μL), concomitant GC dose (mg/day), VDI, and safety. Study 2: The 1-year effectiveness and safety of MPZ (MPZ group, N=15) and the IVCY (IVCY group, N=18) were compared in patients with severe EGPA with organ involvements. The MPZ group was treated with high-dose GC and MPZ, and the IVCY group was treated with high-dose GC and IVCY. The primary endpoint was remission-achieving rate, and secondary endpoints were GC-free remission rate, changes in BVAS, eosinophil counts, concomitant GC dose, VDI, MPZ and IVCY (10-15 mg/kg biweekly, six times) retention rate, and safety.Results:Study 1: The primary endpoint was 72.7% (24/33), and GC-free remission was achieved in 48.5% (16/33). There were no significant changes in BVAS, eosinophil counts, and concomitant GC dose in the Pre-MPZ group (BVAS 0→0, eosinophil count 374→168, GC 6.0→5.0). BVAS, eosinophil counts, and concomitant GC dose significantly decreased in the MPZ group (BVAS 0→0 (p<0.001), eosinophil count 168→32.3 (p=0.002), GC 5.0→0 (p<0.001)). VDI increased significantly in the Pre-MPZ group (3.0→4.0 (P=0.010)), but not in the Post-MPZ group (4.0→4.0 (P=0.083)). The MPZ retention rate was 90.9% (30/33), and adverse events were significantly lower in the Post-MPZ group (p=0.022). Study 2: The two groups had no significant difference in patient background. The primary endpoint was 66.7% (10/15) in the MPZ group and 22.2% (4/18) in the IVCY group, which was significantly higher in the MPZ group (p=0.010). GC-free remission rate was 13.3% (2/15) in the MPZ group and 0% (0/18) in the IVCY group (p=0.110). BVAS decreased significantly in both groups (MPZ group: 16.0 to 0, IVCY group: 17.0 to 0), and there was no significant difference between groups. Eosinophil counts also decreased significantly in both groups (MPZ group: 6992→17.5, IVCY group: 5210→78), with no significant difference between the groups. The concomitant GC dose decreased significantly in both groups (MPZ group: 50→2.5, IVCY group: 58→6.0). The concomitant GC dose (MPZ group vs. IVCY group: 2.5 vs. 6.0 (p<0.001)) and GC reduction rate (%) (MPZ vs. IVCY: -94.0 vs. -86.7 (p<0.001)) were significantly higher in the MPZ group. VDI at 12 months was 1.0 in the MPZ group and 2.0 in the IVCY group, significantly lower in the MPZ group. MPZ retention rate was 100%, and the IVCY completion rate was 72.2%. Adverse events occurred in 26.7% (4/15) in the MPZ group and 61.1% (11/18) in the IVCY group, significantly lower in the MPZ group (p= 0.048). In all 48 patients, the remission-achieving and GC-free remission rates were 70.8% (34/48) and 37.5% (18/48), respectively.Conclusion:The effectiveness and safety of MPZ in the maintenance and remission induction phases were demonstrated. Based on the higher GC-sparing effect, MPZ can be a proper therapeutic option to achieve GC-free remission and avoid irreversible organ damage in real clinical practice.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Masanobu Ueno Masanobu Ueno has received a speaking fee from GlaxoSmithKline., Ippei Miyagawa: None declared, Satoshi Kubo Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca and also research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Yusuke Miyazaki Yusuke Miyazaki has received speaking fees from Eli Lilly and GlaxoSmithKline., Yusuke Miyazaki has received a grant from GlaxoSmithKline., Kentaro Hanami: None declared, Koshiro Sonomoto: None declared, Shingo Nakayamada S. Nakayamada has received speaking fees from Bristol-Myers, AstraZeneca, Pfizer, GlaxoSmithKline, Astellas, Asahi-kasei, Sanofi, Abbvie, Eisai, Chugai, Gilead, and Boehringer Ingelheim., S Nakayamada has received research grants from Mitsubishi-Tanabe., Yoshiya Tanaka Yoshiya Tanaka has received speaking fees from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, Taiho., Yoshiya Tanaka has received grants from Mitsubishi-Tanabe, Eisai, Chugai, Taisho.

Background:Lupus nephritis (LN) is one of the most critical organ involvement in systemic lupus erythematosus (SLE). While there are effective treatment options such as MMF and cyclophosphamide, there are still unmet medical needs: refractory cases to existing therapies and difficult cases to reduce GC. Therefore, the emergence of new treatment options is expected. Recently, rituximab (RTX), an anti-CD20 antibody, is expected to be a new treatment option for LN based on the results of clinical trials. Indeed, the EULAR recommends the use of RTX for refractory LN. In Japan, RTX for LN was approved in August 2023, ahead of the world. However, the effectiveness and safety of RTX in real-world clinical practice have not been fully verified.Objectives:This study aimed to assess the effectiveness and safety of RTX for LN in real-world clinical practice.Methods:This study included 117 patients with LN. The effectiveness and safety were compared between High-dose GC + RTX group (RTX group, n = 58) and high-dose GC + CY and MMF group (standard of care: SoC group, n = 59) in remission induction therapy for patients with LN. Both groups were treated in addition to standard of care including HCQ. Selection bias was minimized by the propensity score-based inverse probability of treatment weighting (PS-IPTW) method and compared between the groups. The primary endpoint was the Complete Renal Response (CRR) achieving rate and uPCR less than 1.0 (g/g・cre) at week 52. Secondary endpoints were treatment retention rate, safety, SLEDAI, and GC reduction rate at 52 weeks. Peripheral blood immunophenotypes (CD4 T cell, CD8 T cell, B cell, NK, monocyte, DC) of LN(n=76) just before the remission induction therapy were compared to those of age- and sex-matched HC (n=109). Additionally, the impact of RTX on immunophenotypes was evaluated by comparing before and after RTX in 21 patients.Results:The 52-week treatment retention rate was 94.8% (55/58) in the RTX group and 83.1% (49/59) in the SoC group, with no difference between the two groups. Infusion reactions were the most frequent adverse event in the RTX group at 20.7% (12/58), and infections were the most frequent in the SoC group at 47.5% (28/59). SLEDAI scores significantly decreased in both groups. Patient baseline characteristics were adjusted by PS-IPTW method. The primary endpoint achievement rate was 1) CRR achieving rate: RTX group; 58.1%, SoC group; 52.2% (p=0.38), uPCR<1.0 achieving rate: RTX group; 83.2%, SoC; 81.3% (p=0.54). There was no significant difference between the groups. There was also no difference in GC sparing effect, the secondary endpoint, between the two groups (RTX group; -81.1%, SoC group; -89.3%, p=0.65). Immune phenotyping revealed increased class-switched memory B cells (CM B cells: CD19+ CD27+ IgM-) and plasmocytes (CD19+ CD27+ CD38+) in SLE compared to HCs. Naïve B cells (CD19+ CD27- IgM+), CM B cells, and plasmocytes disappeared after 26 weeks of RTX introduction. plasmocytes remained low for 52 weeks in cases that achieved the primary endpoint(n=16). On the contrary, CM B cells and plasmocytes increased in cases that did not achieve the primary endpoint (n=5). Patients who relapsed within 3 years after achieving the primary endpoint by RTX were 5/16 cases, in which CM B cells and plasmocytes increased again. Naïve B cells re-elevated while CM B cells and plasmocytes remained absent in the non-relapse cases.Conclusion:Rituximab can be an effective treatment option in real-world clinical practice achieving remission and suppressing relapse by depleting CM B cells and plasmocytes.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Masanobu Ueno Masanobu Ueno has received a speaking fee from GlaxoSmithKline., Shingo Nakayamada S. Nakayamada has received consulting fees, speaking fees, and/or honoraria from Bristol-Myers, AstraZeneca, Pfizer, GlaxoSmithKline, Astellas, Asahi-kasei, Sanofi, Abbvie, Eisai, Chugai, Gilead, and Boehringer Ingelheim, and has received research grants from Mitsubishi-Tanabe., S. Nakayamada has received consulting fees, speaking fees, and/or honoraria from Bristol-Myers, AstraZeneca, Pfizer, GlaxoSmithKline, Astellas, Asahi-kasei, Sanofi, Abbvie, Eisai, Chugai, Gilead, and Boehringer Ingelheim, and has received research grants from Mitsubishi-Tanabe., Ippei Miyagawa: None declared, Satoshi Kubo Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca and also research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Yusuke Miyazaki Y. Miyazaki has received speaking fees from Eli Lilly, and has received research grants from GlaxoSmithKline., Kentaro Hanami: None declared, Koshiro Sonomoto: None declared, Hiroaki Tanaka: None declared, Yoshiya Tanaka Yoshiya Tanaka has received speaking fee from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, and Taiho., Yoshiya Tanaka has received grant from Mitsubishi-Tanabe, Eisai, Chugai, and Taisho.

Background:Systemic lupus erythematosus (SLE) is characterized by a wide diversity of clinical manifestations, and the etiology may vary among patients. Immunophenotyping of peripheral blood has revealed dysregulation in the differentiation of B cells compared to that in healthy individuals. However, the correlation between these differences and clinical symptoms remains unclear. Moreover, the appropriate sub-grouping of lupus patients holds the potential to pave the way for precision medicine.Objectives:The purpose of this study was to identify peripheral blood immunological abnormalities in SLE, assess their diversity, and attempt to subgroup lupus patients. Additionally, we investigated the relationship between these subgroups and clinical symptoms as well as prognosis.Methods:Immunophenotyping of immunocompetent cells in peripheral blood, encompassing T cells, B cells, NK cells, dendritic cells, and monocytes, was conducted using multicolor flow cytometry following the antibody set proposed by NIH/FOCIS. The analysis included 123 untreated patients with SLE and 75 healthy subjects. The immunophenotypes were assessed as a percentage within peripheral blood mononuclear cells. The outcomes underwent cluster analysis, employing both the Ward’s method with Euclidean distance to stratify the patient population. Based on the tree diagram, the number of clusters was set to four. Visualization of the clusters was performed using Uniform Manifold Approximation and Projection (UMAP) method. Clinical symptoms and recurrence rates at 3 years were evaluated within each subgroup.Results:All patients in the study were untreated, with a median age of 42 years. The median disease duration was 5 months. The disease activity, as measured by SLEDAI, averaged 11, and 66% of the patients had either one BILAG A or two Bs (BILAG A1 or B2). Overall, lupus patients exhibited an elevation in double-negative B cells (1.4% vs 0.6%) and plasmablasts (1.4% vs 0.2%) in peripheral blood compared to healthy controls. In T cells, there was an elevation in activated CD4 (1.8% vs 0.6%), activated CD8 (4.7% vs 0.6%), and activated Th1 cells (0.8% vs 0.3%). Additionally, there was a reduction in CD16+ NK cells (5.6% vs 10.3%). Cluster analysis based on this immunophenotype stratified untreated SLE patients into four major groups and visualized them using UMAP (Figure 1). One of the four clusters exhibited an almost identical immunophenotype to that of healthy controls and comprised the smallest number of patients (cluster 1). The remaining three clusters were nearly evenly distributed, distinguished by a relatively low proportion of plasmablasts with a notable increase in double-negative B cells (cluster 2), a tenfold increase in plasmablasts (cluster 3), and an elevation in activated regulatory T cells (cluster 4). Cluster 2 was the youngest in terms of age, with no differences observed in disease duration or gender. In these four clusters, cluster 1 exhibited the lowest disease activity, with BILAG A1 or B2 at 40%, whereas cluster 3, characterized by markedly elevated plasmablasts, showed high disease activity, with BILAG A1 or B2 at 88%, and hypocomplementemia was also prevalent. An analysis of the 3-year clinical course of patients undergoing high-dose glucocorticoid therapy revealed that cluster 1 did not experience a single relapse, while cluster 3 had more than 20% relapses. The remaining clusters exhibited approximately 10% relapses.Conclusion:Immunophenotyping facilitated the stratification of untreated lupus patients into four distinct groups. While SLE is characterized by an elevation of double-negative B cells and plasmablasts, these immune cell subsets exhibited differential behavior within the SLE population. Cluster characterized by a significant increase in plasmablasts exhibited notably high disease activity and relapse rates.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca, has received research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Shingo Nakayamada has received speaking fees from Bristol-Myers, UCB, Astellas, Abbvie, Eisai, Pfizer, and Takeda, has received research grants from Mitsubishi-Tanabe, Novartis, and MSD, Yusuke Miyazaki has received epeaking fees from Astra-Zeneca, speaking fees, and/or honoraria from Eli Lilly and GlaxoSmithKline, Yasuyuki Todoroki: None declared, Masanobu Ueno: None declared, Katsuhide Kusaka: None declared, Satsuki Matsunaga: None declared, Hiroaki Tanaka: None declared, Naoaki Ohkubo: None declared, Hidenori Sakai: None declared, Ippei Miyagawa: None declared, Kentaro Hanami: None declared, Yoshiya Tanaka has received speaking fees and/or honoraria from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, Taiho, has received research grants from Mitsubishi-Tanabe, Eisai, Chugai, Taisho.

Background:It is unclear as to which SLE patients respond favorably to belimumab (BEL), and the impact of such treatment on peripheral blood immunophenotype in this population remains unknown.Objectives:This study aimed to clarify peripheral immunophenotype of patients with SLE who successfully discontinued glucocorticoids (GC) by intervention with BEL in the maintenance phase.Methods:Patients with SLE (n=146), who were in the maintenance therapy phase with a SELENA-SLEDAI score of less than 10 and receiving glucocorticoid therapy at a prednisolone-equivalent dose of 0.2 mg/kg/day or less, were assessed. They were divided into the standard of care (SoC) group (46 patients who received hydroxychloroquine or mycophenolate mofetil) and the BEL group (100 patients who received BEL with SoC). The efficacy of BEL group was compared with SoC group after adjustment by propensity score-based inverse probability of treatment weighting (PS-IPTW). Based on the standard human immune cell subset classification protocol by NIH/FOCIS, peripheral immunophenotypes were analyzed in BEL group and SoC group, and were compared.Results:After PS-IPTW adjustment, no differences in patient characteristics were shown between the SoC and BEL groups. The retention rate of BEL at 52w was 98.0%. SELENA-SLEDAI scores improved after 52w in both groups. The BEL group also had significantly lower GC doses at 52w (p=0.0028) and 31.8% of the BEL group successfully discontinued GC, whereas on 2.1% of the SoC group did (p=0.0043). The incidence of infections was significantly lower in the BEL group compared to the SoC group before PS-IPTW (BEL, 4.0% vs. SoC, 17.5%, p=0.089). The baseline peripheral immunophenotypes were similar between the two groups. In the BEL group, the proportion of activated T follicular helper cells (p=0.0073), IgD-CD27-B (DNB) cells (p=0.0088) and plasmocytes (p=0.0092) decreased significantly at 26w. There were no significant changes in the SoC group. At 26 weeks, the proportion of DNB cells (p=0.0328) and plasmocytes (p=0.0415) was significantly lower in patients who discontinued glucocorticoids (GCs) compared to those who were unable to discontinue them. Multiple logistic regression analysis showed that GC discontinuation was associated with low GC doses, low SLEDAI scores at BEL initiation, decreased IgG levels at 52w, and a low percentage of DNB cells and plasmocytes at 26w.Of those who discontinued GCs in the BEL group, 81.3% (6/26) did not experience a flare-up of SLE one year after discontinuation. In peripheral blood immunophenotyping six months after discontinuation of GC, the proportion of DN Bcells and plasmocytes was increased in patients who relapsed after discontinuation of GC. On the other hand, the proportion of DN Bcells and plasmocytes did not change in those who did not relapse after discontinuation of GC.Conclusion:Intervention with BEL in patients with SLE reduced DNB cells and plasmocytes, thereby controlling the disease activity and enabling GC discontinuation. Among patients who received low GC doses and had low SLEDAI scores, those with decreased IgG levels at 52w successfully discontinued GCs.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Yusuke Miyazaki Y. Miyazaki has received lecture fees from AstraZeneca, GlaxoSmithKline, Astellas, Eli Lilly., Shingo Nakayamada: None declared, Satoshi Kubo Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca and also research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Hiroaki Tanaka: None declared, Kentaro Hanami: None declared, Shunsuke Fukuyo: None declared, Ippei Miyagawa: None declared, Yasuyuki Todoroki: None declared, Yoshino Inoue: None declared, Yurie Satoh-Kanda: None declared, Masanobu Ueno: None declared, Yoshiya Tanaka Y. Tanaka has received Speakers bureau from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, Taiho., Y. Tanaka has received Grant/research support from Mitsubishi-Tanabe, Eisai, Chugai, Taisho.

Background:Rituximab and tocilizumab exhibit potential as molecular targeted therapies for skin fibrosis in patients with systemic sclerosis. While rituximab has demonstrated improvement in skin fibrosis in a limited number of double-blind studies, tocilizumab has shown a favorable trend in a larger double-blind study. However, there is insufficient data on the real-world effectiveness of both drugs in daily clinical practice, and prognostic factors remain unclear.Objectives:To assess and compare the efficacy of rituximab and tocilizumab in treating skin fibrosis relative to standard of care (SoC), while also investigating the factors influencing the effectiveness of each treatment.Methods:In a comparative analysis, 32 patients treated with rituximab alongside SoC, 29 patients receiving tocilizumab with SoC, and 32 patients in the SoC-only group were examined. To mitigate potential selection bias in patient backgrounds across the three groups, propensity score-based inverse probability of treatment weighting was applied. The primary endpoint was the alteration in the modified Rodnan skin score (mRSS) after 24 weeks. Additionally, flow cytometric immune cell profiling, known as the “Human Immunology Project” by NIH/FOCIS, was conducted, and microvascular damages were assessed using nailfold video capillaroscopy. As an exploratory endpoint, we examined peripheral blood immunophenotypic characteristics and the extent of microvascular damages in cases exhibiting improved skin fibrosis.Results:The mRSS values were 14.0, 13.1, and 13.5 for the rituximab, tocilizumab, and SoC groups, respectively, with no significant differences among the three groups. Other parameters such as disease duration (7.2, 8.1, and 7.9 years) and age (59.9, 60.7, and 60.2 years) also exhibited no notable differences. After 6 months of treatment, the improvement in skin score was -3.6 points for rituximab, -2.1 points for tocilizumab, and +0.9 points for SoC. Both rituximab and tocilizumab demonstrated significant improvement compared to SoC (p < 0.001, p = 0.005), with no significant difference between rituximab and tocilizumab (p = 0.24) (Figure 1). The analysis of factors contributing to the effectiveness of each drug revealed that neither rituximab nor tocilizumab exhibited a clinical background predictive of efficacy. Peripheral blood immunophenotyping revealed associations between the percentages of effector memory CD4+ T cells, central memory CD4+ T cells, Th1, Th17, plasmablasts, and CD16+ NK cells and improved skin fibrosis at 24 weeks with rituximab. In multivariate analysis, only plasmablasts were inversely correlated with the efficacy of rituximab (p = 0.003) (Table 1). Conversely, no immune cells contributed to predicting the effectiveness of tocilizumab. Nailfold capillaries analysis showed that tocilizumab tended to improve cases with normal or giant capillaries, while those with hemorrhage or loss of capillaries were less likely to improve (p = 0.20).Conclusion:Both rituximab and tocilizumab exhibited improvement in skin fibrosis compared to SoC. For rituximab, the immunophenotype may be useful in treatment selection. However, there is no identified indicator for tocilizumab, necessitating further biomarker studies.Table 1.Pearson correlation coefficientunivariate analysismultivariate analysisCD4+ T cellsNaive0.0240.895Central memory-0.4020.020.982Effector memory-0.3750.0310.207TEMRA0.1630.365Activated-0.1360.451CD8+ T cellsNaive0.2530.156Central memory-0.2180.224Effector memory0.0080.967TEMRA0.0360.841Activated0.0990.584CD4+ T cell subsetsTh1-0.3580.0410.352Activated Th1-0.130.471Th17-0.3920.0240.063Activated Th170.0490.785Tfh0.0180.923Activated Tfh-0.0190.916Naive Treg0.0850.637Memory Treg0.0020.991Activated Treg0.0940.601B cellsNaive-0.3170.073IgM memory0.0050.976Class-switched memory-0.1050.562Double negative-0.2980.092Plasmablasts0.5460.0010.003NK cellsCD16+ NK cell0.3580.0410.954CD16- NK cell-0.2760.119MonocytesClassical0.2740.122Non classical0.3260.064Dendritic cellsMyeloid0.1160.522Plasmacytoid0.0980.589Figure 1.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca., has received research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Yurie Satoh-Kanda: None declared, Yasuyuki Todoroki: None declared, Ryuichiro Kanda: None declared, Hiroaki Tanaka: None declared, Masanobu Ueno: None declared, Yoshino Inoue: None declared, Yusuke Miyazaki has received speaking fees from Eli Lilly and GlaxoSmithKline., Ippei Miyagawa: None declared, Kentaro Hanami: None declared, Shingo Nakayamada has received speaking fees from Bristol-Myers, UCB, Astellas, Abbvie, Eisai, Pfizer, and Takeda, has received research grants from Mitsubishi-Tanabe, Novartis, and MSD., Yoshiya Tanaka has received speaking fees from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, Taiho., has received research grants from Mitsubishi-Tanabe, Eisai, Chugai, Taisho.

Background:Achievement and maintenance of the Lupus Low Disease Activity State (LLDAS) are necessary for the long-term prevention of organ damage progression in patients with systemic lupus erythematosus (SLE); however, minor flares lower the achievement and maintenance rates, and consequent treatment changes such as glucocorticoid (GC) dose increase. Therefore, there is a demand for therapeutic strategies to control disease activity and achieve dose reduction or discontinuation of GCs using molecular targeted drugs.Objectives:This study aimed to analyze the safety and efficacy of anifrolumab in patients with SLE who experience minor flares after achieving LLDAS in real-world clinical practice.Methods:In this retrospective observational study, we assessed 65 SLE patients who experienced minor flares after achieving LLDAS. Of these, 30 were treated with the addition of glucocorticoids (GCs) or immunosuppressants, forming the standard of care (SoC) group. The remaining 35 patients were treated with the additon of only anifrolumab, constituting the anifrolumab group. Minor flare was defined as the revised Safety of Estrogens in Lupus Erythematosus National Assessment Flare Index. The LLDAS achievement rate at 26 weeks in the SoC and anifrolumab group were compared after adjusting with inverse probability of treatment weighting using propensity score (PS-IPTW).Results:The retention rate of anifrolumab was 97.1% (34/35 patients) at week 26. No significant difference was observed in the patient background between the two groups after adjustment by PS-IPTW. There was no difference between the two groups in either the LLDAS achievement rate or the DORIS remission rate at week 12 after the onset of minor flares followed by treatment intensification. At week 26, the anifrolumab group had a higher the rate of LLDAS achievement (SoC: anifrolumab=53.3:85.7%, p=0.0042) and DORIS remission (SoC: anifrolumab=6.7:25.7 %, p=0.0412). The SoC group did not exhibit a significant decrease in GC dose at 26 weeks, while the anifrolumab group exhibited a significant decrease. GC doses at week 26 were lower in the anifrolumab group (SoC: anifrolumab=6.9±3.5: 3.1±3.8mg/day, prednisolone equivalent, p<0.0001). The incidence of adverse events were fewer in the anifrolumab group (p=0.0012), especially infections (p=0.0169).Compared with patients treated for minor flares by GC dose increase (GC dose increase group, n=18) after adjustment by PS-IPTW, the anifrolumab group had a higher rate of LLDAS achievement (GC dose increase: anifrolumab=39.8:89.1%, p<0.0001) and DORIS remission (GC dose increase: anifrolumab=12.4:40.8%, p=0.0011). GC dose at week 26 was lower in the anifrolumab group (GC dose increase: anifrolumab=7.2±2.6:3.1±3.8 mg/day, prednisolone equivalent, p=0.0001). In anifrolumab group, three cases discontinued GC 26weeks after introduction of anifrolumab. The incidence of adverse events (p=0.0086) and infection (p=0.0442) were significant lower in the anifrolumab group.Conclusion:The current study demonstrated the safety and efficacy of anifrolumab in patients with minor flares who once achieved LLDAS but had difficulty maintaining LLDAS. These findings suggested that disease activity could be improved by initiating anifrolumab therapy alone without GC dose increase in patients with minor flares. Anifrolumab may prevent the accumulation of disease-induced organ damage caused by SLE and drug-induced organ damage caused by GCs, thereby improving long-term QOL and prognosis in patients with SLE.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of Interests:Yusuke Miyazaki Y.Miyazaki has received speaking feesfrom Bristol-Myers, Pfizer, GlaxoSmithKline, Astellas, Asahi-Kasei, and Boehringer Ingelheim., Shingo Nakayamada: None declared, Satoshi Kubo Satoshi Kubo has received speaking fees from Eli Lilly, GlaxoSmithKline, Bristol-Myers, Abbvie, Eisai, Pfizer, Astra-Zeneca. research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Satoshi Kubo has received research grants from Daiichi-Sankyo, Abbvie, Boehringer Ingelheim, and Astellas., Satsuki Matsunaga: None declared, Hiroaki Tanaka: None declared, Kentaro Hanami: None declared, Shunsuke Fukuyo: None declared, Yurie Satoh-Kanda: None declared, Yoshino Inoue: None declared, Yasuyuki Todoroki: None declared, Masanobu Ueno: None declared, Yoshiya Tanaka Y. Tanaka has received Speakers bureau from Eli Lilly, AstraZeneca, Abbvie, Gilead, Chugai, Behringer-Ingelheim, GlaxoSmithKline, Eisai, Taisho, Bristol-Myers, Pfizer, Taiho, Y. Tanaka has received Grant/research support from Mitsubishi-Tanabe, Eisai, Chugai, Taisho.

Tofacitinib (TOFA) and baricitinib (BARI) have been widely used in many regions for treatment of rheumatoid arthritis (RA). The selection of JAK inhibitor for RA treatment based on patient type remains a major concern. The differences of efficacy between each Janus kinase (JAK) inhibitors have not been clarified in the patients with RA in clinical practice. Here, we compared the efficacy between TOFA and BARI in clinical practice. A retrospective observational study. The efficacy of TOFA (n=156) in patients with RA was compared with BARI (n=138). Selection bias was reduced to a minimum using propensity score-based inverse probability of treatment weighting (IPTW). We analyzed the trajectories of changes in disease activity in patients receiving TOFA or BARI using growth mixture modeling (GMM). Multivariable logistic regression analysis was performed to identify factors contributing to belonging to treatment-resistance group defined by GMM. The observation period of the study was 24 weeks. No significant difference was observed in patient characteristics between the TOFA and BARI groups in after adjustment by propensity score-based IPTW. The retention rates over 24 weeks did not differ between the TOFA and BARI groups. No difference was observed in the incidence of adverse events in the TOFA and BARI groups. Clinical disease activity index (CDAI) at week 24 after the introduction of JAK inhibitors was 8.0 ± 8.9 and 6.2 ± 7.2 in the TOFA and BARI group, respectively. The rates of CDAI-remission at week 24 in the TOFA and BARI groups were 43/153 (28.3%) and 57/141 (40.4%), respectively. Compared to the TOFA group, the BARI group showed a significantly lower CDAI (⊿CDAI=-1.9, 95% confidence interval: -3.7 to -0.3, p=0.02) and a significantly higher rate of CDAI-remission (odds ratio: 1.7, 95% CI: 1.1–2.7, p=0.04) at week 24. Similarly, at week 24, SDAI was significantly lower in the BARI group (TOFA vs. BARI = 10.1 ± 9.9 vs. 7.3 ± 7.5, ⊿SDAI=-2.2, 95% CI: -4.2 to -0.2, p=0.04), and the rates of SDAI-remission (OR: 1.6, 95% CI: 1.0–2.6, p=0.04). The patients were divided into two groups: patients with MDA to HDA at baseline (Group 1) and patients with HDA at baseline than Group 1 (Groups 2 and 3) based on the analysis of the trajectories of CDAI using GMM, In Groups 1 and 2, disease activity was improved immediately after the introduction of JAK inhibitors. In Group 3, disease activity was partially improved, and LDA was not achieved at week 24 after the introduction of JAK inhibitors. The patients in Group 3 were resistant to treatment (Group 3: treatment-resistance group). When multivariable logistic regression analysis was performed for all patients receiving JAK inhibitors, the factors contributing to belonging to treatment-resistance group were: high baseline HAQ-DI score (OR: 1.76, 95% CI: 1.09–2.84, p=0.02) and high number of biological disease-modifying anti-rheumatic drugs (bDMARDs) used before JAK inhibitors (OR: 1.51, 95% CI: 1.16–1.95, p=0.002) and TOFA use (OR: 2.13, 95% CI: 1.05–4.30, p=0.03). Next, multivariable logistic regression analysis was separately performed for each treatment group. The patients receiving more bDMARDs before the JAK inhibitor were more likely to belong to treatment-resistance group in the TOFA group (OR: 1.76, 95% CI: 1.24–4.06). Among patients with RA who received TOFA, those who had received ≥4 bDMARDs before the introduction of TOFA were more likely to be classified into the treatment-resistant group. In the BARI group, multivariable logistic regression analysis did not identify any factors associated with belonging to treatment-resistance group. TOFA may be partially effective in patients resistant to many bDMARDs. Consequently, efficacy may differ between TOFA and BARI. Because TOFA was less effective in RA patients resistant to ≥4 bDMARDs, the present study suggests that BARI may be more appropriate for RA patients resistant to many bDMARDs. Yusuke Miyazaki Speakers bureau: Eli Lilly, Shingo Nakayamada Speakers bureau: Bristol-Myers, UCB, Astellas, Abbvie, Eisai, Pfizer, Takeda, Kazuhisa Nakano Speakers bureau: Bristol-Myers, Sanofi, AbbVie, Eisai, Eli Lilly, Chugai, Pfizer, Takeda, and Mitsubishi-Tanabe, Satoshi Kubo Speakers bureau: Bristol-Myers, Yoshino Inoue: None declared, Yoshihisa Fujino: None declared, Yoshiya Tanaka Speakers bureau: Abbvie, Daiichi-Sankyo, Chugai, Takeda, Mitsubishi-Tanabe, Bristol-Myers, Astellas, Eisai, Janssen, Pfizer, Asahi-kasei, Eli Lilly, GlaxoSmithKline, UCB, Teijin, MSD, and Santen