J Korean Med Sci.  2008 Jun;23(3):514-520. 10.3346/jkms.2008.23.3.514.

Mechanism of Humoral and Cellular Immune Modulation Provided by Porcine Sertoli Cells

• Lee HM
• Oh BC
• Lim DP
• Lee DS
• Lim HG
• Park CS
• Lee JR

• KMID: 1786893
• DOI: http://doi.org/10.3346/jkms.2008.23.3.514

Abstract

The understanding of main mechanisms that determine the ability of immune privilege related to Sertoli cells (SCs) will provide clues for promoting a local tolerogenic environment. In this study, we evaluated the property of humoral and cellular immune response modulation provided by porcine SCs. Porcine SCs were resistant to human antibody and complement-mediated formation of the membrane attack complex (38.41+/-2.77% vs. 55.02+/-5.44%, p=0.027) and cell lysis (42.95+/-1.75% vs. 87.99 +/-2.25%, p<0.001) compared to immortalized aortic endothelial cells, suggesting that porcine SCs are able to escape cellular lysis associated with complement activation by producing one or more immunoprotective factors that may be capable of inhibiting membrane attack complex formation. On the other hand, porcine SCs and their culture supernatant suppressed the up-regulation of CD40 expression (p<0.05) on DCs in the presence of LPS stimulation. These novel findings, as we know, suggest that immune modulatory effects of porcine SCs in the presence of other antigen can be obtained from the first step of antigen presentation. These might open optimistic perspectives for the use of porcine SCs in tolerance induction eliminating the need for chronic immunosuppressive drugs.

Keyword

Porcine Sertoli Cells; Immune Privilege; Humoral Immune Response; Cellular Immune Response; Tolerance

MeSH Terms

Animals
Antibodies, Heterophile/immunology
Antibody Formation/*immunology
Antigens, CD40/immunology
Aorta/cytology
Cell Line, Transformed
Cell Survival/immunology
Complement Membrane Attack Complex/immunology
Complement System Proteins/immunology
Dendritic Cells/cytology/immunology
Endothelial Cells/cytology/immunology
Epitopes/immunology
Humans
Immune Tolerance/*immunology
Immunity, Cellular/*immunology
Male
Mice
Mice, Inbred C57BL
Sertoli Cells/cytology/*immunology
Swine
*Tissue Engineering
Transplantation, Heterologous

Figure

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  Fig. 1 Expression of Gal-α1,3-Gal (Gal) on immortalized porcine Sertoli (NPSCi) or aortic endothelial (PECi) cells. Cells cultured as a monolayer on chamber slides and fixed for 2 hr with absolute cold ethanol were immunostained for IB4 lectin (brown). Most of NPSCi (A) and PECi (B) cells are positive to the Gal epitope. FACS analysis of non-fixed NPSCi (C) and PECi (D) cells stained with FITC-conjugated IB4 lectin (1:50) for 1 hr to detect Gal (solid line). Also, there are no significant differences in the binding of human IgM to binding of human xenoreactive antibodies to NPSCi (E, 76.95±6.03%) or PECi (F, 80.76±2.44%) cells (p>0.05).

• 

  Fig. 2 Formation of membrane attack complex (MAC) on the immortalized porcine Sertoli (NPSCi, A, 38.41±2.77%) and aortic endothelial (PECi, B, 55.02±5.44%) cells in the presence of normal human serum. Both cell lines show the significant difference in the formation of MAC (p=0.027). Gray histograms correspond to the control cells that were not pre-incubated with human serum, and solid histograms represent specific staining of human C5b-9. The data are presented as mean±SD of three individual experiments in triplicate.

• 

  Fig. 3 Susceptibility of immortalized porcine Sertoli (NPSCi) or aortic endothelial (PECi) cells to human natural antibody- and complement-mediated lysis. Negative control values (media alone) for NPSCi and PECi cells were arbitrarily set equal to 100%, and the relative percentage of cell viability was calculated for each condition. The data are presented as the mean±SD of three individual experiments in triplicate. Means bearing different superscript letters are significantly different at p<0.05. M, media alone; M+C, media+rabbit complement; iNHS, heat-inactivated normal human serum; NSH, normal human serum.

• 

  Fig. 4 The effects of immortalized porcine Sertoli cells on the activation of dendritic cells. Immortalized porcine SCs (NPSCi cells) do not activate DCs (empty bar) but inhibit cell surface expression of CD40 in the presence of LPS. The data are presented as the mean±SD of three individual experiments in triplicate. Means bearing different superscript letters are significantly different at p<0.05. MFI, mean fluorescence index; ciDC, immature DC differentiated in the presence of Dexa and D3; iDC, immature DC; LPS, lipopolysaccharide.

• 

  Fig. 5 The effects of culture supernatant of immortalized porcine Sertoli cells (iSPNT) on the activation of dendritic cells (DCs) in the presence of LPS. CD40 expression on DCs was significantly inhibited in the presence of iSPNT. But iSPNT did not affect the expression of CD80 and I-Ab molecules on the surface of LPS stimulated DCs. The data are presented as the mean±SD of three individual experiments in triplicate. Means bearing different superscript letters are significantly different at p<0.05. MFI, mean fluorescence index; iDC, immature DC; LPS, lipopolysaccharide.

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