Histo-Immunological Detection of Collagen Fractions

Mancini, R. E.; Paz, Mercedes A.; Vilar, O.; Davidson, O. W.; Barquet, J.

doi:10.3181/00379727-118-29838

Cited by 5 publications

(2 citation statements)

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“…In respect to the immunofluorescent staining of basement membranes our results are at variance with several reports which demonstrated stain ing of glomerular basement membranes with antisera to skin or tendon collagen [2,[7][8][9], but they are in agreement with the observations of Mancini et al [4] who did not find such a reaction. Our observations, however, were only made with purified antibodis since each original an tiserum exhibited strong reactivity with kidney glomeruli.…”

Section: Discussioncontrasting

confidence: 57%

“…The demonstration of considerable amounts of antibodies to impurities, like serum proteins [10] or acidic connective tissue components [11,12], in 'anti-collagen' sera -even when highly purified collagen was used as im munogen -has, however, shed some doubt on the reliability of such re sults. Contradictory findings on the immunofluorescent staining of glome rular basement membranes by antibodies obtained to either rat, human or chicken tendon collagen [2,4,[7][8][9] seem to emphasize the possibility of interference by antibodies to noncollageneous antigens.…”

Section: Introductionmentioning

confidence: 99%

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Purified Antibodies to Collagen: An Immunofluorescence Study of Their Reaction with Tissue Collagen

Wick¹,

Furthmayr²,

Timpl³

1975

Int Arch Allergy Immunol

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Antibodies to soluble calf or rat skin collagen were purified by appropriate immunoadsorption and separated into three subfractions directed either to antigenic sites on unfolded α-chains (denatured collagen), to nonhelical sites, or to helical sites exposed on the triple helical molecule. In indirect immunofluorescence tests each of the antibody solutions reacted with collagen of skin and kidney tissue, although the latter two antibody solutions appeared to be more active. Distinct activity was also observed with antibodies to the N-terminal antigenic determinant of rat skin collagen α₂-chain, a structure usually involved in cross-linking. Indirect immunofluorescence tests with anti-collagen sera on sections of skin resulted in the staining of the whole dermis, while anti-procollagen sera revealed binding only to the uppermost subepithelial layer of the dermis (stratum papillare). On kidney sections only the interstitial connective tissue reacted with purified anti-collagen or anti-procollagen sera. Both skin and glomerular basement membranes remained unstained with either kind of purified antibodies. However, antisera not subjected to immunoadsorption do react with the glomerular basement membrane. Antibodies to noncollagenous contaminants are considered to be responsible for this finding which emphasizes the necessity to use purified antibodies exclusively for this type of immunofluorescence analysis.

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Section: Discussioncontrasting

confidence: 57%