Fitch J M, Gross J, Mayne R, Johnson-Wint B, Linsenmayer T F
Proc Natl Acad Sci U S A. 1984 May;81(9):2791-5. doi: 10.1073/pnas.81.9.2791.
To determine whether type V collagen is antigenically masked in situ by its fibrillar organization, two different methods were used to perturb selectively the structure of collagen fibrils in sections of embryonic chicken corneas. The experimentally modified tissues were probed by immunohistochemical procedures with monoclonal antibodies against types V and I. A lathyritic agent was used to block crosslinking of newly synthesized collagen. This results in reversible temperature-sensitive alterations in fibrillar packing, such that freshly formed collagen fibrils retain their aggregated state at 37 degrees C but become dissociated upon cooling. Type V-specific immunofluorescence remained masked at 37 degrees C but was revealed at 0 degree C. The effect of temperature was partially reversible, indicating that type V collagen is normally unavailable for antibody binding because of its fibrillar arrangement. In sections of normal corneas, treatment with corneal collagenase, which degrades type I collagen, but not type V, also unmasked the latter. This implicates type I collagen as the masking agent. We propose that collagen types I and V are incorporated together in heterotypic fibrils.
为了确定V型胶原在原位是否因其纤维组织而被抗原性掩盖，我们使用了两种不同的方法来选择性地扰乱胚胎鸡角膜切片中胶原纤维的结构。通过免疫组织化学方法，用抗V型和I型单克隆抗体对经实验修饰的组织进行检测。一种致畸形药物被用于阻断新合成胶原的交联。这导致纤维堆积出现可逆的温度敏感性改变，使得新形成的胶原纤维在37℃时保持聚集状态，但在冷却时会解离。V型特异性免疫荧光在37℃时仍被掩盖，但在0℃时显现出来。温度的影响部分可逆，这表明V型胶原由于其纤维排列，通常无法与抗体结合。在正常角膜切片中，用角膜胶原酶处理，该酶可降解I型胶原而非V型胶原，也能使后者暴露出来。这表明I型胶原是掩盖剂。我们提出I型和V型胶原共同整合到异型纤维中。
