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ÛÚÙ   «“√ “√°“√·æ∑¬å·ºπ‰∑¬·≈–°“√·æ∑¬å∑“߇≈◊Õ°                                 ªï∑’Ë ˆ©∫—∫∑’Ë Û°—𬓬π-∏—𫓧¡ ÚııÒ



                 °“√«‘‡§√“–Àåª√‘¡“≥°√¥¬Ÿ‚√𑧠                  ·≈–¡’§“√å∫Õπ‰¥ÕÕ°‰´¥å ı% ‡ªìπ‡«≈“ Ù ™—Ë«‚¡ß.

                    «‘‡§√“–ÀåÀ“ª√‘¡“≥°√¥¬Ÿ‚√π‘§„πµ—«Õ¬à“߇ÀÁ¥À≈‘π®◊Õ    ¿“¬À≈—ß°“√‡æ“–‡≈’È¬ß π”µ—«Õ¬à“߇≈◊Õ¥∑’Ë∂Ÿ°°√–µÿâπ
                                              ÒÒ
             ‚¥¬«‘∏’ meta-hydroxydiphenyl method .              ı ‰¡‚§√≈‘µ√ ¡“¬âÕ¡¥â«¬‚¡‚π‚§≈π—≈·Õπµ‘∫Õ¥’¬åµ‘¥
                 °“√«‘‡§√“–Àå™π‘¥πÈ”µ“≈‡™‘߇¥’ˬ«‚¥¬«‘∏’∑”ªØ‘°‘√‘¬“∑“ß  ©≈“°¥â«¬ “√‡√◊Õß· ß™π‘¥µà“ß Ê (Becton Dickinson, San

             ‡§¡’·≈–«‘∏’ Thin layer chromatography              Jose, CA) Û ™π‘¥ §◊Õ CD3 (∑’≈‘¡‚ø‰´µå) ∑’˵‘¥©≈“°¥â«¬
                    «‘‡§√“–Àå™π‘¥πÈ”µ“≈‡™‘߇¥’ˬ«„πµ—«Õ¬à“߇ÀÁ¥À≈‘π®◊Õ   “√‡√◊Õß· ß fluorescein isothiocyanate (FITC), CD69
                                    ÒÚ
             ‚¥¬«‘∏’ acid hydrolysis/TLC .                      (early avtivation marker) ∑’˵‘¥©≈“°¥â«¬ “√‡√◊Õß· ß phy-
                 °“√‡µ√’¬¡µ—«Õ¬à“߇æ◊ËÕ∑¥ Õ∫                    coerythrin (PE) ·≈– CD45 (‡¡Á¥‡≈◊Õ¥¢“«) ∑’˵‘¥©≈“°
                    ≈–≈“¬ “√ °—¥πÈ” ·≈– “√ °—¥·Õ≈°ÕŒÕ≈å „π      ¥â«¬ “√‡√◊Õß· ß peridinin chlorophyll protein (PerCP)

             phosphate buffered saline (PBS).  à«π “√ °—¥∑’ˉ¡à  ‡ªìπ‡«≈“ Òı π“∑’, ·≈â«∑”„À⇡Á¥‡≈◊Õ¥·¥ß·µ°¥â«¬ “√≈–≈“¬
             ≈–≈“¬πÈ” ≈–≈“¬„π DMSO Ú% „ππÈ”‡≈’Ȭ߇´≈≈å RPMI1640.  lysing. ®“°π—Èπ®÷ߪíòπ≈â“ߥ⫬ wash buffer (2% fetal calf
                 °“√µÕ∫ πÕߢÕß≈‘¡‚ø‰´µåµàÕ “√ °—¥‡ÀÁ¥À≈‘π®◊Õ    serum „π PBS) ‡µ‘¡æ“√“øÕ√å¡—≈¥’‰Œ¥å Ò% ≈ß„πµ–°Õπ‡´≈≈å.

                    ‡®“–‡≈◊Õ¥®“°Õ“ “ ¡—§√„ à≈ß„πÀ≈Õ¥∑’Ë¡’ “√°—π  ®“°π—Èπ®÷ß𔉪«‘‡§√“–Àåº≈¥â«¬‡§√◊ËÕß‚ø≈‰´‚µ¡‘‡µÕ√å
             ‡≈◊Õ¥·¢Á߇Œæ“√‘π. ®“°π—Èπ„™âµ—«Õ¬à“߇≈◊Õ¥ª√‘¡“≥ ı  FACSort (Becton Dickinson) §à“√âÕ¬≈–¢Õß∑’≈‘¡‚ø‰´µå∑’Ë

             ‰¡‚§√≈‘µ√ °√–µÿâπ¥â«¬ “√ °—¥µà“ßÊ ®“°‡ÀÁ¥À≈‘π®◊Õ∑’Ë  ¡’  CD69 ª√“°ØÕ¬Ÿà∫πº‘«À≈—ß®“°∂Ÿ°°√–µÿâπ «‘‡§√“–Àå‚¥¬
             §«“¡‡¢â¡ Ò ‰¡‚§√°√—¡/¡‘≈≈‘≈‘µ√ À√◊Õ°√–µÿâπ√à«¡°—∫ Sta-  „™â‚ª√·°√¡ CellQuest (Becton Dickinson) ¥—ß· ¥ß„π
             phylococcal enterotoxin B (SEB) (Sigma, St.Louis, MO)  √Ÿª∑’Ë Ò.

             ª√‘¡“≥ ı ‰¡‚§√°√—¡ ‚¥¬¡’ SEB ‡ªìπµ—«§«∫§ÿ¡º≈∫«°         °“√«‘‡§√“–Àåº≈°“√∑¥≈Õß
             ·≈–πÈ”‡≈’Ȭ߇´≈≈å RPMI1640 ‡ªìπµ—«§«∫§ÿ¡º≈≈∫. º ¡„Àâ       «‘‡§√“–À姫“¡·µ°µà“ߢÕß§à“‡©≈’ˬ√–À«à“ßÀ≈Õ¥
             ‡¢â“°—π·≈–𔉪‡æ“–‡≈’Ȭ߄πµŸâÕ∫∑’ËÕÿ≥À¿Ÿ¡‘ Û˜ Õß»“‡´≈‡´’¬   §«∫§ÿ¡·≈–À≈Õ¥∑’Ë∂Ÿ°°√–µÿâπ¥â«¬ “√ °—¥®“°‡ÀÁ¥À≈‘π®◊Õ



































             √Ÿª∑’Ë Ò °“√«‘‡§√“–À凴≈≈å≈‘¡‚ø‰´µå¥â«¬‡§√◊ËÕß‚ø≈‰´‚µ¡‘‡µÕ√å ‡¡◊ËÕ¬âÕ¡¥â«¬‚¡‚π‚§≈π—≈·Õπµ‘∫Õ¥’¬åµàÕ CD3/CD69/CD45. ‡´≈≈å≈‘¡‚ø‰´µå
                    “¡“√∂·¬°‰¥â‚¥¬¥Ÿ SSC/CD45 (R1) (´â“¬¡◊Õ) ·≈–§à“√âÕ¬≈–°“√· ¥ßÕÕ°¢Õß CD69 ∫π∑’≈‘¡‚ø‰´µå  “¡“√∂«‘‡§√“–À剥â
                   ®“° quadrant ∫π¢«“ (¢«“¡◊Õ)
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