Characterization of Cryoglobulins by Immunoblotting

Musset, Lucile; Diemert, Marie‐Claude; Taibi, F; Dû, Lê Thi Huong; Cacoub, Patrice; Léger, J.-M.; Boissy, G; Gaillard, O; Galli, Jonathan

doi:10.1093/clinchem/38.6.798

Cited by 198 publications

(55 citation statements)

References 7 publications

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“…There are, however, unusual cryoglobulins whose immunochemical structure cannot be fitted into these three classes. MC formed by oligoclonal IgM and traces of polyclonal IgG, for example, have been described and their inclusion in the Brouet classification as a type II-type III variant has been proposed [10]. This IgM microheterogeneity has been seen as indirect evidence of a possible transition from type III to type II.…”

Section: Classification Of Cryoglobulinsmentioning

confidence: 99%

The cryoglobulins: an overview

Dammacco

Sansonno

Piccoli

et al. 2001

Eur J Clin Investigation

210

156

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Cryoglobulins are cold-precipitable immunoglobulins associated with a number of infectious, autoimmune and neoplastic disorders. Their appearance along with rheumatoid factor (RF) can be considered a normal event in the clearance of immune complexes and rarely produces any symptoms. The association between hepatitis C virus (HCV) and mixed cryoglobulinemia (MC) has been rendered evident since the recognition of serological markers of HCV infection. There is thus every reason to suppose that direct or indirect involvement of B cells on the part of the HCV results in their persistent stimulation, clonal expansion and release of molecules with RF activity. The formation of RF/IgG immune complexes is the key pathogenetic mechanism. The close correlation between HCV infection and MC also throws new light on the interpretation of autoimmune phenomena in the course of viral infection and on the close link between autoimmune diseases and lymphoproliferative disorders. The higher risk of non-Hodgkin's lymphoma (NHL) displayed by HCV positive subjects, especially in the Mediterranean basin, suggests that the HCV's chronic lymphoproliferative drive may progress towards frank lymphoid neoplasia. The presence of MC does not represent an in situ or`occult' NHL, because recent evidences indicate that none of the clones interpreted as predominant displays the molecular features of a true neoplastic process. The cryoglobulinemic syndrome is probably the consequence of pathogenic noxae that act upon the immune system of a host in which regulation of the peripheral T cell response appears to be in some way altered.

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Section: Classification Of Cryoglobulinsmentioning

confidence: 99%

The cryoglobulins: an overview

Dammacco

Sansonno

Piccoli

et al. 2001

Eur J Clin Investigation

210

156

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“…Cryoglobulins were isolated from the patients' sera, purified and characterized by immunoblotting at 37°C as previously described [21]. In the present study, patients were considered to have a significant cryoglobulin if they had a minimum serum cryoglobulin level of 0.05 g/L at two determinations.…”

Section: Detection and Characterization Of Cryoglobulinsmentioning

confidence: 99%

Lower expression of CD81 B‐cell receptor in lymphoproliferative diseases associated with hepatitis C virus infection

Cacoub¹,

Bourlière

Hausfater³

et al. 2003

Journal of Viral Hepatitis

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Chronic hepatitis C virus (HCV) infection is frequently associated with type II mixed cryoglobulinaemia (MC), a benign lymphoproliferative disease (LPD). More recently, HCV has been implicated as a possible aetiologic factor of B-cell non-Hodgkin lymphoma (B-NHL). CD81, a B-cell surface receptor, has been proposed as a receptor for HCV binding and entry in circulating B cells. The stimulation of CD81 complex enables B cells to respond to lower concentrations of antigen and finally induces B-cell proliferation. We studied the phenotypic expression of CD81, CD19 and CD5 on circulating B cells in HCV patients LPD-positive or LPD-negative. Sixty-two patients were anti-HCV antibody positive. Among HCV positive patients, 44 were HCV RNA positive with an histologically proven chronic active hepatitis of whom 10 had a B-NHL, 14 an MC and 24 no extrahepatic manifestation. Eighteen patients were HCV RNA negative with evidence of resolved infection. A control group included 40 healthy subjects. Peripheral blood mononuclear cells (PBMC) were stained for surface expression of CD81, CD19 and CD5 using monoclonal antibodies, and were analyzed by flow cytometry. The percentage of PBMC expressing CD81, CD19 and CD5 receptors were compared between the groups by univariate analysis. Logistic regression model variables were then evaluated to correlate the presence of an LPD with HCV infection characteristics (i.e. age, gender, genotype, duration of infection, HCV RNA positivity, liver histological lesions), or phenotypic expression of CD81, CD19 and CD5 receptors on PBMC. HCV antibody-positive compared with HCV-negative subjects had a higher expression of CD19 receptor (23 +/- 13 vs 13 +/- 1%, P = 0.003). Among HCV RNA positive-patients, LPD+ compared with LPD- patients had a lower expression of CD81 (58 +/- 28 vs 82 +/- 18%, P = 0.001) and CD5 receptor (66 +/- 16 vs 74 +/- 13%, P = 0.04). In multivariate analysis, the expression of CD81 receptor was a negative (OR = 0.15, 95% CI = 0.04-0.64, P = 0.01) and CD19 receptor a positive (OR = 4.81, 95% CI =1.29-17.88, P = 0.02) predictive factor for an LPD. We found two negative predictive factors for HCV RNA positivity, i.e. age (OR = 0.23, 95% CI. = 0.08-0.62, P = 0.003) and the expression of CD81 receptor (OR = 0.34, 95% CI = 0.13-0.89, P = 0.02). In patients with a chronic active HCV infection, the presence of a lymphoproliferative disease, either MC or B-NHL, is associated with lower expression of CD81 and higher expression of CD19 receptor on peripheral B cells.

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“…The immunoglobulin composition of the purified Cg was estimated by immunofixation (Paragon Beckmann) and immunoelectrophoresis using poly-and monospecific antisera [3]. Protein concentration in Cg was estimated by diluting aliquots of Cg in 0´1 m NaOH and reading the absorbance in a spectrophotometer at 280 nm [22]. The values were derived from a standard curve using purified human gamma globulin (Polyglobin N R from Bayer, Germany).…”

Section: Preparation and Classification Of Cryoglobulinsmentioning

confidence: 99%

Occurrence of C-reactive protein in cryoglobulins

Weiner

Prasauskas²,

Lebrecht

et al. 2001

Clinical and Experimental Immunology

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SUMMARYA previous case report described the formation of a complex between a monoclonal IgA with cryolabile properties and C-reactive protein (CRP). Our study provides the first evidence for the frequent occurrence of CRP in cryoglobulins (Cg) of all three types according to Brouet's classification. We performed a systematic immunochemical analysis of cryoglobulins from 18 patients by Western blotting and in 15 of 18 cryoprecipitates a single band (23 KD), immunoreactive with anti-CRP antibody, was demonstrable irrespective of the clonal composition of the cryoglobulins. This band was detectable in 4/5 of type I, in 6/8 of type II, and in 5/5 of type III cryoprecipitates, classified according to Brouet et al. In addition, the complement proteins C1q and C3 were present in nearly all CRP-containing cryoglobulins, presumably reflecting previous activation of the classical complement pathway at least. All three CRP-negative cryoprecipitates were derived from sera with low cryoglobulin content (1±2 g/l). Longitudinal investigation of 23 cryoprecipitates from seven patients confirmed that successful detection of CRP by Western blotting depends on the protein concentration of the cryoglobulins. Since complexed CRP was previously shown to be an effective activator of complement, via C1q binding, CRP may modulate pathophysiologic effects mediated by cryoglobulins in vivo.

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Characterization of Cryoglobulins by Immunoblotting

Cited by 198 publications

References 7 publications

The cryoglobulins: an overview

The cryoglobulins: an overview

Lower expression of CD81 B‐cell receptor in lymphoproliferative diseases associated with hepatitis C virus infection

Occurrence of C-reactive protein in cryoglobulins

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