We'd like to understand how you use our websites in order to improve them. Register your interest. Immunological and structural studies were performed in 42 cases of cryoglobulinemia. Seven of them were immunochemically characterized as single-type cryoglobulins 3 IgG and 4 IgM , whereas the remaining 35 were mixed IgG-IgM cryoglobulins. Mixed cryoglobulins were divided into 2 groups, depending on the polyclonal 25 cases or monoclonal 10 cases character of the IgM component.
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Mixed cryoglobulinemic vasculitis is associated with monoclonal B cell expansion in patients with chronic hepatitis C HCV infection.
B cell depletion therapy using rituximab, a CD20 monoclonal antibody, has been successful in achieving remission from symptomatic disease. This study investigated whether B cell depletion therapy has an impact on activation of HCV-specific T cell phenotype and function. Using flow cytometry and Enzyme-Linked Immunospot assay, the number of activated and tissue-like B cells and number of T cells expressing Programmed cell death protein 1 PD-1 , T-cell immunoglobulin and mucin-domain containing-3 TIM-3 , and multiple cytokines were measured before and after rituximab therapy.
B cell depletion therapy results in recovery of T cell exhaustion and function in patients with HCV cryoglobulinemic vasculitis. Recent clinical trials have shown HCV-MC vasculitis patients can be effectively treated with rituximab, an anti-CD20 monoclonal antibody, resulting in symptomatic relief. Chronic HCV infection is associated with increases in T cells with an exhausted phenotype defined as cells expressing Programmed cell death protein 1 [PD-1], T-cell immunoglobulin and mucin-domain containing-3 [TIM-3] , the hallmark of which is a lack of polyfunctionality, and is more commonly observed in those cells specifically responding to HCV antigens.
In the present study, we sought to further characterize T cell phenotype and function, and reversal of T cell exhaustion, with respect to HCV specificity before and after rituximab therapy in patients with HCV-MC. The clinical trial primary endpoint was remission 6 months from randomization. After study month 6, patients randomized to the control arm were offered therapy with rituximab if they continued to have active manifestations of vasculitis delayed therapy. This study used 19 patients who received rituximab therapy overall 10 patients from the therapy and 9 patients from the control arm.
All experiments were performed in compliance with relevant laws and institutional guidelines and in accordance with the ethical standards of the Declaration of Helsinki.
Peripheral blood was collected by venipuncture and peripheral blood mononuclear cells PBMCs were isolated from white blood cells by the standard Ficoll-Hypaque Plus Amersham Biosciences, Uppsala, Sweden density gradient separation technique as previously described. All cultures were performed in duplicate. Brefeldin A and monensin were added at 36 hours to block the Golgi apparatus and allow intracellular cytokine accumulation.
Cells were harvested and live, CFSE cells were assessed for phenotype and function using the antibodies described previously. The gating strategy is shown in supplemental Figure 1. All samples were tested in duplicate. We also explored if there was a difference in the change of these parameters between patients who received immediate vs. The clinical and demographic characteristics of patients from the clinical trial, and that rituximab therapy was successful with a clinical response, have been previously described.
There was no difference in the change between patients who received immediate rituximab compared to those who received delayed therapy. There was a significant increase in polyfunctionality more than one cytokine from pre-therapy 1. Consistent with the lowered expression of exhaustive markers, T cells exhibited increased secretory function when stimulated with HCV peptides, suggesting reversal of exhaustion as the mechanism of this phenomenon. The monoclonal expansion of B cells that leads to immune complex mediated small vessel vasculitis also leads to T cell exhaustion.
Rituximab therapy results in rapid depletion of circulating tissue-like memory B cells, and our study showed an increase in HCV-specific T cell function following rituximab therapy. Previously, we described B cell depletion in the same patients result in T cell activation These results further extend these observations suggesting depletion of B cells and reversal of inflammatory response lead to recovery of HCV-specific T cells.
The exact mechanism involved is not well understood, but an indirect effect of B cells that reduce inflammatory response and drive immune exhaustion may be the major driver of the aberrant antiviral response.
These results suggest a specific effect of B cell depletion therapy and remission on HCV specific immunity, which is a unique finding. Our study had limitations. The sample size was small and the follow-up time was limited; long-term follow-up would have allowed us to study T cell functionality with B cell recovery and, sustenance of response, and disease relapse. Second, it is difficult to quantify immunological response, which makes it challenging to evaluate the clinical significance of enhanced antiviral immunity.
Since we did not have long-term follow-up, we were not able to evaluate substance of this response. Other factors that could mediate inflammatory response in these patients such as B lymphocyte stimulator BLyS-1 , interleukin-1 IL-1 , and interleukin 1 receptor antagonist IL-IRA , could also play a role in explaining these findings. In this regard, we have 3 possible explanations.
First, the variability in the T cell frequency in input cells is a confounding factor that could impact the levels of cytokine. Future studies should focus on evaluating B and T cell phenotype and specificity in HCV-MC vasculitis patients treated with DAAs, in order to understand whether recovery of antiviral immunity is associated with enhanced clearance of virus and achievement of SVR.
Supplemental Figure 1. Surface string and intracellular staining after Brefeldin treatment were also performed as described in the Methods. Conflict of interest: None of the authors have any conflicts of interest to report. Experimental Ethics: All experiments were performed in compliance with relevant laws and institutional guidelines and in accordance with the ethical standards of the Declaration of Helsinki.
Informed consent was obtained by the human subjects for this study. Additional Supporting Information may be found in the online version of this article. National Center for Biotechnology Information , U. J Med Virol. Author manuscript; available in PMC May 1. Author information Copyright and License information Disclaimer.
Copyright notice. The publisher's final edited version of this article is available at J Med Virol. Conclusion B cell depletion therapy results in recovery of T cell exhaustion and function in patients with HCV cryoglobulinemic vasculitis.
PBMC collection Peripheral blood was collected by venipuncture and peripheral blood mononuclear cells PBMCs were isolated from white blood cells by the standard Ficoll-Hypaque Plus Amersham Biosciences, Uppsala, Sweden density gradient separation technique as previously described. RESULTS Study Population The clinical and demographic characteristics of patients from the clinical trial, and that rituximab therapy was successful with a clinical response, have been previously described.
Open in a separate window. Figure 1. Figure 2. Figure 3. Figure 4. Click here to view. Footnotes Conflict of interest: None of the authors have any conflicts of interest to report Experimental Ethics: All experiments were performed in compliance with relevant laws and institutional guidelines and in accordance with the ethical standards of the Declaration of Helsinki.
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Only comments seeking to improve the quality and accuracy of information on the Orphanet website are accepted. For all other comments, please send your remarks via contact us. Only comments written in English can be processed. Mixed cryoglobulinemia MC is a rare multisystem disease characterized by the presence of circulating cryoprecipitable immune complexes in the serum, manifested clinically by a classical triad of purpura, weakness and arthralgia. It is considered to be a rare disorder, but its true prevalence remains unknown.
Crioglobulinemia: Un modello di malattia da immunocomplessi. Studi immunochimici e strutturali
Only comments seeking to improve the quality and accuracy of information on the Orphanet website are accepted. For all other comments, please send your remarks via contact us. Only comments written in English can be processed. Type I cryoglobulinemia is frequently asymptomatic per se but patients may develop acrocyanosis, retinal hemorrhage, Raynaud's phenomenon, and arterial thrombosis.
Non-hepatitis C virus crioglobulinemic vasculitis: a case series of nine patients. Boris A. Hybner IV ; Fabiana B. Goulart IV ; Paulo M. Demographics, clinical, laboratory data, treatments and follow-up are reported. In three patients, CV was attributed to connective tissue disease and in one to a monoclonal gammopathy of undetermined significance MGUS. Five patients presented essential cryoglobulinemia.