Paper # 096 | Versión en Español |
Thymic microenvironmental changes in human congenital syphilis
Eliene C. Fonseca, Mônica P. Almeida, Évlin H. Maia, Dora M. F. Menezes and Wilson Savino*
[Title] [Introduction] [Results] [Discussion] [Pictures] [Bibliography]
Thymic tissue were obtained from necropsies of ten stillborn and four neonates dying from congenital syphilis, and two controls with normal thymuses, died due to prematurity and intrauterine anoxy.
HISTOLOGY: Thymus fragments were routinely processed, paraffin embedded and 4-5µm sections were stained with hematoxilin-eosin, Gomori's reticulin and Gomori's trichromic. Picrosyrius stain polarization method was used to differentiate type I collagen (very birrefringent, with thick yellow or red fibers in a dark background) from type III collagen -less refringent, thin greenish fibers (22).
IMMUNOFLUORESCENCE: antibodies specifically recognizing distinct extracellular matrix components were obtained from Pasteur Institute - Lyon, France. All were polyclonal immunosera anti types I, III and IV collagens. Fibronectin, laminin and all other antiserum did not exhibit any cross-reactivity with any other ECM molecule. The goat anti-rabbit Ig antibodies coupled to either fluorescein or rhodamine and fluorescein-labled anti-human IgG and IgM were purchased from Biosys (Coupiegne, France).
IMMUNOPEROXIDASE: Polyclonal antibody anti-human cytokeratin (wide spectrum) immunostained thymic epithelial cells (TEC) network. Goat anti-rabbit-biotinilated antibody and streptavidin-biotin-peroxidase complex were used to perform the immunoreaction (all reagents from Dako Corporation, Carpinteria, USA).
Distribution of Extracellular Matrix componentes in normal human thymus |
|||||
Connective septae | |||||
Thymocytes |
TEC |
Lobular basement membrane |
Septae |
BloodVassel |
|
Type I Col. | - |
- |
± |
+++ |
+++ |
Type III Col. | - |
- |
± |
+ |
+++ |
Type IV Col. | - |
± |
+++ |
++ |
+++ |
Fibronectin | - |
± |
+++ |
++ |
+++ |
Laminin | - |
++ |
+++ |
++ |
+++ |