Karpova Ia D, Lyupina Iu V, Astakhova T M, Stepanova A A, Erokhov P A, Abramova E B, Sharova N P
Bioorg Khim. 2013 Jul-Aug;39(4):400-10. doi: 10.1134/s1068162013040092.
The dynamics of the expression of LMP7 and LMP2 proteasome subunits in embryonic and early postnatal development of rat spleen and liver is investigated in comparison with the dynamics of chymotrypsin-like and caspase-like proteasome activities and expression of MHC (major histocompatibility complex) class I molecules. The immune subunits LMP7 and LMP2 distribution in spleen and liver cells in the development process is also studied. A mutual for both organs tendency to the increase of the expression of both LMP7 subunit and LMP2 one on P21 (the 21st postnatal day) as compared to the embryonic period is discovered. However, the total proteasome level is shown to be constant. At definite development stages, the dynamics of immune subunits expression in the spleen and liver was different. In the spleen gradual enhancement of both immune subunits level being detected on P1, P18 and P21, in the liver gradual enhancement periods on E16 (the 16th embryonic day) and E18 changed to the stage of the shrink of immune subunits level on P5. This level did not reliably change till P18 and was augmented on P21. The alterations revealed were accompanied by chymotrypsin-like activity raise and caspase-like activity drop in spleen by P21 as compared with the embryonic period, which proves the enlargement of proteasome ability to form antigenic epitopes for MHC class I molecules. In the liver, both activities increased by P21 in comparison with the embryonic period. Such dynamics of caspase-like activity can be explained not only by the change of proteolytic constitutive and immune subunits, but also by additional regulatory mechanisms. Besides, it is discovered that the increment of immune subunits expression in the early spleen development is connected with the process of successive forming the white pulp by B- and T-lymphocytes enriched by immune subunits. In the liver, the growth of immune subunits level by P21 was accompanied by their expression expansion in hepatocytes, while their plunge by P5 may be related to the loss of liver function of a primary lymphoid organ of the immune system by this stage and disappearance of B-lymphocytes enriched by immune proteasomes in it. In the spleen and liver, MHC class I molecules were revealed at the periods of the raise of proteasome immune subunits level. On E21 , the liver was enriched by neuronal NO-synthase, its level decreased after birth and enhanced to P18. This fact indicates the possibility of the induction of the immune subunits LMP7 [character: see text] LMP2 expression in hepatocytes in signal way with neuronal NO-synthase participation. The results obtained prove that T-cell immune response with spleen participation as regards rat liver cells is possible starting with P19-P21 stage. First, at this period, white pulp T-area is formed in the spleen. Second, enhanced immune proteasomes and MHC class I molecules levels in hepatocytes can procure antigenic epitopes formation from foreign proteins and their delivery to cell surface for their subsequent presentation for cytotoxic T-lymphocytes.
研究了大鼠脾脏和肝脏在胚胎期和出生后早期发育过程中LMP7和LMP2蛋白酶体亚基的表达动态,并与类胰凝乳蛋白酶样和类半胱天冬酶样蛋白酶体活性以及主要组织相容性复合体(MHC)I类分子的表达动态进行了比较。还研究了发育过程中脾脏和肝脏细胞中免疫亚基LMP7和LMP2的分布。发现与胚胎期相比,两个器官在出生后第21天(P21)时LMP7亚基和LMP2亚基的表达均有增加的共同趋势。然而,蛋白酶体的总水平显示为恒定。在特定的发育阶段,脾脏和肝脏中免疫亚基的表达动态不同。在脾脏中,在P1、P18和P21时检测到两种免疫亚基水平逐渐增强;在肝脏中,在胚胎第16天(E16)和E18时免疫亚基水平逐渐增强,到P5时转变为免疫亚基水平下降阶段。该水平直到P18时没有明显变化,在P21时升高。与胚胎期相比,脾脏中这些变化伴随着到P21时类胰凝乳蛋白酶样活性升高和类半胱天冬酶样活性下降,这证明蛋白酶体形成MHC I类分子抗原表位的能力增强。在肝脏中,与胚胎期相比,到P21时两种活性均增加。类半胱天冬酶样活性的这种动态变化不仅可以用蛋白水解组成性和免疫亚基的变化来解释,还可以用其他调节机制来解释。此外,发现脾脏早期发育过程中免疫亚基表达的增加与富含免疫亚基的B淋巴细胞和T淋巴细胞相继形成白髓的过程有关。在肝脏中,到P21时免疫亚基水平的升高伴随着其在肝细胞中的表达扩展,而到P5时其下降可能与该阶段肝脏作为免疫系统初级淋巴器官功能的丧失以及其中富含免疫蛋白酶体的B淋巴细胞的消失有关。在脾脏和肝脏中,在蛋白酶体免疫亚基水平升高的时期发现了MHC I类分子。在E21时,肝脏中富含神经元型一氧化氮合酶,其水平在出生后下降并在P18时升高。这一事实表明,在神经元型一氧化氮合酶参与下,肝细胞中可能以信号方式诱导免疫亚基LMP7[特征:见正文]LMP2的表达。获得的结果证明,从P19 - P21阶段开始,大鼠肝细胞的T细胞免疫反应可能涉及脾脏。首先,在这个时期,脾脏中形成了白髓T区。其次,肝细胞中增强的免疫蛋白酶体和MHC I类分子水平可以从外来蛋白质中获得抗原表位的形成,并将其递送到细胞表面,以便随后呈递给细胞毒性T淋巴细胞。