Detection of pemphigus vulgaris antigen on COLO and SCaBER tumor cell lines by the immunoblot technique

Nm, Mirza; Mohimen, A; Ar, Ahmed

doi:10.1007/bf00372096

Cited by 8 publications

(4 citation statements)

References 20 publications

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“…This sensitive modified IB assay demonstrated negative results with normal human serum and sera from other autoimmune disease. Our present study demonstrated a 130 kDa band with normal human skin and a 105 kDa band with COLO-16 lysate as expected [13] with sera of PV patients. Positive control sera from patients with BP reacted with 230 kDa and 160 kDa bands with both lysates as expected [8].…”

Section: Discussionsupporting

confidence: 55%

“…SDS-PAGE was performed by Laemmli's method [12] using a 7.5% running gel and a 3% stacking gel, pH 6.8, with a discontinuous buffer system as described previously [13]. Separated proteins were electrophoretically transferred to a PVDF protein sequencing membrane (BR, Richmond, Calif.) according to the method described by Towbiu et al [14].…”

Section: Immunoblottingmentioning

confidence: 99%

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Comparison of an indirect immunofluorescence assay and a modified sensitive immunoblot assay for the study of the autoantibody in pemphigus vulgaris

Mohimen

Ahmed

1995

Arch Dermatol Res

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Some patients with pemphigus vulgaris (PV) have positive direct immunofluorescence (DIF) but are negative by indirect immunofluorescence (IIF). The purpose of this study was (1) to compare the sensitivity of an IIF assay with an immunoblot (IB) assay, (2) to compare the IIF and the IB assay in PV patients in whom the clinical picture and DIF were consistent, but the IIF was negative and (3) to compare the IIF and the IB assay in patients in clinical remission for 3 years or more. A comparison was made of the titers of PV autoantibody in the IIF assay using monkey esophagus as substrate and the modified sensitive IB assay using preabsorbed normal human skin lysate and COLO-16 lysate as a substrate in the three groups of patients. The sensitivity of the Western blot was enhanced by modifications in the extraction procedure of the lysate, by absorption of lysate with normal human serum and by the use of an enzygraphic web. In group 1, comprising 23 PV patients with active generalized disease, the titers of the autoantibody in the IB assay were 2-4-fold higher than in the IIF assay. This difference was highly significant (P = 0.0001). In group 2, comprising 10 patients with limited or minimal PV who were positive on DIF and negative on IIF, all the patients were positive in the IB assay. In group 3, comprising 9 patients clinically free of disease and off all therapy for at least 3 years and negative in IIF assay, all the patients were positive in the IB assay.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionsupporting

confidence: 55%

Section: Immunoblottingmentioning

confidence: 99%

Comparison of an indirect immunofluorescence assay and a modified sensitive immunoblot assay for the study of the autoantibody in pemphigus vulgaris

Mohimen

Ahmed

1995

Arch Dermatol Res

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“…Immunoblots were produced by incubating subject serum with detergent-solubilized normal skin cells or squamous carcinoma cells after electrophoresis in polyacrylamide gel in the presence of SDS. Two modifications of the method described previously (20) were introduced to increase sensitivity. Cell extracts were passed through a Sepharose 4B column previously coupled to normal human serum to avoid nonspecific binding by serum proteins and transferred after electrophoresis to PVDF membranes (Bio-Rad Laboratories, Richmond, CA).…”

Section: Methodsmentioning

confidence: 99%

“…Serum samples from the study and control subjects were tested for antibody to the 130-kD (keratinocytes) (4) and 105-kD (COLO 16 tumor cell line) (19) antigens characteristic of PV by a Western blot method (20). Immunoblots were produced by incubating subject serum with detergent-solubilized normal skin cells or squamous carcinoma cells after electrophoresis in polyacrylamide gel in the presence of SDS.…”

Section: Methodsmentioning

confidence: 99%

Linkage of pemphigus vulgaris antibody to the major histocompatibility complex in healthy relatives of patients.

Mohimen

et al. 1993

The Journal of Experimental Medicine

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SummaryPemphigus vulgaris (PV) is an autoimmune disease caused by high concentrations of antibody to an epidermal cadherin. The disease is associated with two kinds of HLA-DR4, DQ8 haplotypes dominantly distributed among Jewish patients, and these plus DR6, DQ5 haplotypes in nonJewish patients. Low levels of the PV antibody were found in 48% of a total of 120 asymptomatic parents, children, and siblings of 31 patients, thus exhibiting dominant inheritance. The inheritance of these low levels of antibody in asymptomatic relatives was linked to the major histocompatibihty complex with a highly significant logarithm of the odds score of 9.07, almost always to a DR4 or DR6 haplotype of the patient. Disease appears to occur in susceptible individuals with low levels of antibody when a second factor, either environmental or genetic, induces high levels, sufficient to produce blisters.

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Immunoblot assay to study the pemphigus vulgaris antigen in normal human epidermis and COLO-16 cell line

Mohimen

Ahmed

1993

Arch Dermatol Res

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Detection of pemphigus vulgaris antigen on COLO and SCaBER tumor cell lines by the immunoblot technique

Cited by 8 publications

References 20 publications

Comparison of an indirect immunofluorescence assay and a modified sensitive immunoblot assay for the study of the autoantibody in pemphigus vulgaris

Comparison of an indirect immunofluorescence assay and a modified sensitive immunoblot assay for the study of the autoantibody in pemphigus vulgaris

Linkage of pemphigus vulgaris antibody to the major histocompatibility complex in healthy relatives of patients.

Immunoblot assay to study the pemphigus vulgaris antigen in normal human epidermis and COLO-16 cell line

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