Àΰ£ Á¦´ëÇ÷ÀÇ CD14+ ´ÜÇÙ±¸ ¼¼Æ÷ ¹× CD34+ Á¶Ç÷¸ð¼¼Æ÷¿¡¼ À¯·¡µÈ ¼öÁö»ó¼¼Æ÷ÀÇ Æ¯¼º ºñ±³
Comparison of characteristics of Dendritic Cells derived from CD 14+ Monocyte and CD34+ Hematopoietic Stem Cell in Human Umbilical Cord Blood
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Abstract
¸ñÀû: Á¦´ëÇ÷ÀÇ CD14+ ´ÜÇÙ±¸ ¹× CD34+ Á¶Ç÷¸ð¼¼Æ÷¸¦ ÀÌ¿ëÇÏ¿© µ¿ÀÏÇÑ ¹è¾çÁ¶°Ç¿¡¼ ¼öÁö»ó¼¼Æ÷·Î ºÐȽÃÄÑ ¾òÀº °¢°¢ ÀÇ ¼öÁö»ó¼¼Æ÷¸¦ ÇüÅÂÇÐÀû, Ç¥ÇöÇüÀû ¹× ±â´ÉÀûÀ¸·Î ºñ±³ ºÐ¼®ÇÏ¿© ±× Ư¼ºÀ» ¾Ë°í ¼öÁö»ó¼¼Æ÷¸¦ ÀÌ¿ëÇÑ Ç׾ϸ鿪ġ·á ¿¡¼ ¾î¶² ¼¼Æ÷°¡ ÀûÇÕÇÑ Àü±¸¼¼Æ÷ÀÎÁö ÆÇ´ÜÇϰíÀÚ ÇÑ´Ù.
¿¬±¸ ¹æ¹ý: Á¤»óÀûÀÎ ¸¸»è »ê¸ð¸¦ ´ë»óÀ¸·Î ÇÏ¿© ŹÝÀÌ ¸¸Ã⠵DZâ Àü äÃëµÈ Á¦´ëÇ÷¿¡¼ CD14+ ´ÜÇÙ±¸ (n=17)¿Í CD34+ Á¶Ç÷¸ð¼¼Æ÷ (n=15)¸¦ ºÐ¸®ÇÏ¿© GM-CSF (800 U/mL)°ú IL-4 (500 U/mL)¸¦ ÀÌ¿ëÇÏ¿© ¼öÁö»ó¼¼Æ÷·Î ºÐȽÃŲ ÈÄ LPS (1 Og/mL) ¹× IFN-¥ã (1000 U/mL)¸¦ ÀÌ¿ëÇÏ¿© ¼º¼÷½ÃÄÑ confocal microscopy¸¦ ÀÌ¿ëÇÏ¿© ÇüÅÂÇÐÀû Ư¼ºÀ» ºñ±³ÇÏ¿´À¸¸ç FACS¸¦ ÀÌ¿ëÇÏ¿© Ç¥ÇöÇüÀ» ºÐ¼®ÇÏ¿´´Ù. ELISA kit¸¦ ÀÌ¿ëÇÏ¿© ¼öÁö»ó¼¼Æ÷¿¡¼ »ý»êµÈ IL-12p70ÀÇ ¾çÀ» ºñ±³ÇÏ¿´À¸¸ç È¥ÇÕ¸²ÇÁ±¸ ¹ÝÀÀÀ» ÅëÇØ T cell¿¡ ´ëÇÑ Áõ½Ä´ÉÀ» È®ÀÎÇÏ¿´´Ù.
°á°ú: ´ÜÇÙ±¸ ¹× Áٱ⼼Æ÷¿¡¼ ºÐÈµÈ 8Àϰ ¼º¼÷ ¼öÁö»ó¼¼Æ÷´Â ÇüÅÂÇÐÀûÀ¸·Î µ¿ÀÏÇÑ ¸ð¾çÀ» º¸¿´À¸¸ç CD80, CD83 ¹× CD86À» ÀÌ¿ëÇÑ Ç¥ÇöÇü °Ë»ç¿¡¼µµ À¯»çÇÑ ¹ßÇö Á¤µµ¸¦ º¸¿´´Ù (P>0.05). ±â´ÉÀûÀÎ ºñ±³¸¦ À§ÇÑ È¥ÇÕ ¸²ÇÁ±¸ ¹ÝÀÀ¿¡¼µµ ¸ðµÎ °·ÂÇÑ ¸²ÇÁ±¸ ÀÚ±Ø ÀÛ¿ëÀ» º¸¿´À¸³ª ´ÜÇÙ±¸ ±â¿øÀÇ ¼öÁö»ó¼¼Æ÷´Â 1:20ÀÇ ºñÀ²¿¡¼ °¡Àå °·ÂÇÑ È¿°ú¸¦ º¸ÀÎ ¹Ý¸é Á¶Ç÷¸ð¼¼Æ÷ ±â¿øÀÇ ¼öÁö»ó¼¼Æ÷´Â 1:2ÀÇ ºñÀ²¿¡¼ °¡Àå °·ÂÇÑ È¿°ú¸¦ º¸ÀÌ¸ç ¸ðµç ºñÀ²ÀÇ È¥ÇÕ¸²ÇÁ±¸ ¹ÝÀÀ°Ë»ç¿¡¼ ´Ü ÇÙ±¸ ±â¿øÀÇ ¼öÁö»ó¼¼Æ÷¿¡ ºñÇØ »ó´ëÀûÀ¸·Î À¯ÀÇÇÏ°Ô ³ôÀº Áõ½Ä´ÉÀ» º¸¿´´Ù (P<0.05). IL-12p70 Á¤·®°Ë»ç¿¡¼ µÎ Á¾·ùÀÇ ¼öÁö»ó¼¼Æ÷ ¸ðµÎ 6ÀϰÀÇ ¹Ì¼º¼÷¼¼Æ÷¿¡¼º¸´Ù 8ÀϰÀÇ ¼º¼÷¼¼Æ÷¿¡¼ À¯ÀÇÇÏ°Ô ³ôÀº ¼öÄ¡¸¦ º¸¿´´Ù (P<0.05). ¶ÇÇÑ Á¶Ç÷ ¸ð¼¼Æ÷ ±â¿øÀÇ ¼öÁö»ó¼¼Æ÷¿¡¼ ´ÜÇÙ±¸ À¯·¡ ¼öÁö»ó¼¼Æ÷¿¡ ºñÇØ Åë°èÇÐÀûÀ¸·Î ³ôÀº ¼öÄ¡ÀÇ IL-12p70ÀÇ »ý»ê·®À» º¸¿´´Ù(P<0.05).
°á·Ð: ÀÌ»óÀÇ °á°úµéÀ» °í·ÁÇϸé CD34+ Á¶Ç÷¸ð¼¼Æ÷¿¡¼ ±â¿øµÈ ¼öÁö»ó¼¼Æ÷°¡ Ç׾ϸ鿪¿ä¹ý¿¡ ÀûÇÕÇÑ ¼¼Æ÷ÀÏ ¼ö ÀÖÀ¸³ª µÎ ¼¼Æ÷ ¸ðµÎ È¿°úÀûÀÌ°í ¼º¼÷ÇÑ ¼öÁö»ó¼¼Æ÷·Î ºÐ鵃 ¼ö ÀÖ¾î Ç׾ϸ鿪ġ·á¿¡ À¯¿ëÇÏ°Ô ÀÌ¿ëµÉ ¼ö ÀÖÀ¸¸ç ÀûÀýÇÑ Àü±¸ ¼¼Æ÷ÀÇ ¼±ÅÃÀº Á¦´ëÇ÷¿¡¼ ¾òÀ» ¼ö ÀÖ´Â ¼¼Æ÷ÀÇ Ã¤Ãë È¿¿ë¼º¿¡ Á¿ìµÉ °ÍÀ¸·Î »ý°¢µÈ´Ù.
Objective: To compare the morphological, phenotypical, and functional characteristics of dendritic cells (DCs) generated from two precursor cell sources (CD 14+ monocyte and CD34+ hematopoietic stem cell) of human umbilical cord blood (UCB) under identical ex vivo generation conditions and to determine the best cellular source for DCs-based anticancer immunotherapy.
Methods: CD14+ monocytes and CD34+ hematopoietic stem cells were isolated from human UCB and induced to differentiate into DCs under identical culture conditions using granulocyte macrophage colony stimulating factor (800 U/mL) and IL-4 (500 U/mL). Then maturation of DCs was induced with IFN-¥ã (1000 U/mL) and LPS (1 Og/mL). Morphology was compared with confocal microscopy and phenotypical analysis was done with FACS. The level of IL-12p70 production was determined with ELISA kit and T cell proliferation capacity was assessed with mixed lymphocyte reaction (MLR).
Results: Eight-day-old mature DCs from CD14+ monocytes or CD34+ hematopoietic stem cells had identical morphology. Flow cytometric analysis revealed that CD14+ monocyte-derived DCs (M-DCs) and CD34+ hematopoietic stem cell-derived DCs (CD34-DCs) showed similar enhanced expression of CD80, CD83, and CD86 (P>0.05). The level of IL-12p70 production was significantly higher in mature DCs (8-day-old) than immature DCs (6-day-old) of either source (P<0.05), but it was significantly highly elevated in CD34-DCs (P<0.05). DCs of both precursors showed potent stimulation capacity in MLR, with CD34-DCs having a maximum effect at 1:2 ratio and M-DCs at 1:20 ratio, although CD34-DCs had significantly greater proliferative effects at all ratios (P<0.05).
Conclusion: These findings suggest that CD34-DCs may be a more attractive source for DC-based immunotherapy. But because of morphologic, phenotypical, and functional similarities between the two cellular sources, both are good sources for generating DCs and one is not superior to the other. Thus cellular availability could be an important factor in determining DC-generation protocol to be used if all other issues are equal. Further comparative testing for these two precursors is in need.
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Á¦´ëÇ÷;´ÜÇÙ±¸;Á¶Ç÷¸ð¼¼Æ÷;¼öÁö»ó¼¼Æ÷;Ç׾ϸ鿪ġ·á;Umbilical cord blood;Monocyte;Hematopoietic stem cell;Dendritic cell;Anticancer immunotherapy
KMID :
0358420060490112316
DOI :
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