Distribution of T- & B-cell Series and Macrophages in the Peripheral Blood and the Utero-placental Interface of Pregnant Mice / 대한해부학회지

The experiment was performed to investigate changes of maternal immune status during the pregnancy. We observed the distribution of several immune cells [macrophage, activated B-cell, IgM+ B-cell, Lyt-2+ T-cell and L3T4+ T-cells] in the utero-placental interface and the peripheral blood of Balb/c mice. The experimental animals were divided into seven groups by the gestational ages ; virgin, 2nd, 5th, 8th, 10th, 14th and 19th day of pregnancy. In the utero-placental interface, the distribution patterns of the lymphocytes [both T and B] and macrophages were observed. Histochemical staining by naphthol-AS-MX phosphate sodium salt was used for the detection of activated B-cells. For the detection of macrophage, plasma cell, suppressor cell and helper cell, all the prepared samples reacted with Rat anti-mouse Mac-1, goat anti-mouse IgM, rat anti-Lyt2 and rat anti-L3T4 antibody first, and washed. Second reaction was done with biotinylated anti-rat or anti-mouse IgG anti-bodies, and washed. Avidin-biotin -peroxidase complex and 3, 3`-diamino-benzidine[DAB] were used for the visualization of specific cells. T-cells and B-cells were not observed during the all stages of pregnancy. By contrast, macrophages were observed a few at the perimetrium on the second day of gestation, and they were found at the outermost portion of the trophoblastic layer on the 8th day, and they were observed at the decidua basalis in late pregnancy after the 10th day when the placenta were well developed. In the peripheral blood, activated B-cells were not observed throughout the pregnancy. On the 8th day, the proportion of plasma cells to total mononuclear cells was decreased significantly to 16+/-2.4% compared with the virgin group[22+/-2.6%][p<0.01]. It increased again and it reached 42+/-5.8% on the 14th day and 37+/-4.9% on the 19th day. Helper T-cells were decreased on the 14th day[30+/-2.4%] compared with the normal control[47+/-5.1%]. But, Suppressor T-cells were increased on the 8th day[35+/-2.9%] and the 10th day[33+/-3.6%] compared with the normal controls[27+/-2.3%]. This decrement returned to the level of the normal controls on the 14th day and 19th day. Together with our previous data, we could find the synchronized changes of immune cells in utero-placental interface, uterus draining lymph nodes, peripheral blood and spleen. Therefore, we suggest that macrophages in utero-placental interface may play an important role for the immune responses against the fetal transplantation antigen.

Characterization of Changes in Cells and Extracellular Matrix Components during the Regenerative Processes in the Resected Rat Thyroid / 대한해부학회지

Thyroid gland is composed of follicles surrounded by basement membrane, which is known to be related to the regenerative processes in several organs. This study was performed to observe the morphological details and changes of the basement membrane components, which are the presumtive factors in thyroid regeneration, with time sequences after partial thyroidectomy and then the relations between the components and thyroid regeneration were confirmed. Thyroid galnds of adult male rats were examined before and 1, 2, 3, 4, 5, 7 and 10 days after bilateral partial[50-60%] thyroidectomy. The basememnt membranes of follicles were prominent with high eosinophilicity at later stage when H & E stained. But in PAS and EM staining, the basement membrane of the operated groups didn`t show any specific changes comparing with control group but the dense collagenous fibers at the interfollicular space outside the membrane. Immunostaining of proliferating cell nuclear antigen[PCNA], which reflects the growth fraction, showed 2.3% positivity of total follicular cells in control group. The PCNA positive ratio in experimental groups were 10.3%, 15.9%, 19.9%, 12.2%, 10.5%, 1.9%, 2.7% on postoperative 1, 2, 3, 4, 5, 7, 10 days respectively. In unilaterally thyroidectomized rats, the resected lobe revealed 18.2% of PCNA positive ratio 3 days after operation, while that in the unresected lobe was only 5.9%. This suggested the predominence of local factors in thyroid regeneration. Meanwhile, with the double immunostaining of calcitonin and PCNA discriminate between follicular cells and C-cells among the PCNA positive cells, PCNA positive C-cells were very rare. The immunostaining of laminin, fibronectin, and collagen IV showed varying intensity with the progress of regeneration. Stainability for collagen IV was maintained in all groups with some increase in 5- and 7-day groups. Laminin was well localized to the basement membrane of the control follicle, and the staining intensity for laminin was markedly increased with additional cytoplasmic staining in follicular cells in 1- to 3-day groups and then decreased to attain the control level in 10-day group. In case of fibronectin, the control follicle showed scarce staining, but the resected group showed strong positivity for fibronectin with the most intense staining in 1- to 3-day groups. Immunostaining for fibronectin still showes in 10-day group with the gradual disappearance of immunoreactivity in the center of the lobe. This study revealed the accompaning changes of the basement membrane components during the regeneration process of partially resected thyroid glands. Among them, collagen IV seems to support the structural integrity of the basement membrane. Laminin and fibronectin were supposed to be related to the thyroid follicular regeneration.

Distribution of the Basement Membrane Components in the Rat Thyroid Gland during the pre - and postnatal Period / 대한해부학회지

It is well known that the spatial patterns of basement membrane[BM] components show a close correlation with the morphogenetic process in developing organs during pre- and postnatal period. But little is known about the chronological changes in BM components during the development of thyroid gland. To investigate the correlation of the distribution of BM components with the development of the thyroid gland, immunohistochemical localization of the BM components was performed, using rat fetuses[from 15 to 21 days of gestation] and pups[from 1 to 14 days]. According to the histological features with H.E. staining, prenatal development[El5-21] of the rat thyroid could be divided into 3 representative stages : 1] Undifferentiated primordium of thyroid epithelia, forming cell cords or cell nests [until El5 day]. 2] An intervening stage of cell proliferation with the pattern of lobulation[El6-17 days]. 3] Stage of maturing follicles[El8-21 days]. It could be concluded that differentiation of the thyroid follicle are terminated during the prenatal period and growth in size and number occurs during the first two weeks after birth. By the immunohistochemical localization of laminin[LM], fibronectin[FN], type IV collagen[CIV], the basement membranes investing thyroid follicles could be clearly delineated at 15 days of gestation. Reactivity for LN and CIV associated with BM increased with advancing development, while that of FH was apparently weakened during the 2 weeks after birth. On the other hand immunoreactivity for CIV in the same age group increased, compared to that of previous swage of development. These results indicate that BM components were expressed in distinct spatial patterns with advancing development of thyroid gland. Based on these observations, it could be concluded that these BM components are important determinants in epithelial growth and differentiation during thyroid development.