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Experimental Oncology 22, 99-109, 2000 (September) 99 REVIEW MODERN TRENDS IN CANCER VACCINOTHERAPY V.A. Shlyakhovenko* R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine, Kyiv 03022, Ukraine ÑÎÂÐÅÌÅÍÍÛÅ ÏÎÄÕÎÄÛ Ê ÑÎÇÄÀÍÈÞ ÏÐÎÒÈÂÎÎÏÓÕÎËÅÂÛÕ ÂÀÊÖÈÍ Â.À. Øëÿõîâåíêî* Èíñòèòóò ýêñïåðèìåíòàëüíîé ïàòîëîãèè, îíêîëîãèè è ðàäèîáèîëîãèè èì. Ð.Å. Êàâåöêîãî ÍÀÍ Óêðàèíû, Êèåâ, Óêðàèíà Modern data on cancer vaccines have been reviewed. The mechanism facilitating tumor cells to escape the immune surveillance have been considered with particular emphasis on the significance of tumor-associated (TAA) and tumor-specific (TSS) antigens, proteins of the major histocompatibility complex (MHC), adhesion molecules, and molecules enhancing immune response. The efforts on the elaboration of the recombinant vaccines as well as vaccines on the basis of nucleic acids and glycopeptides have been analyzed. The outlooks for new vectors designed for introducing genetic material into tumor cell genome as well as advantages of liposomes and substances enhancing immune responce have been outlined as well. Recent results and further prospects in the field of cancer vaccinotherapy have been analyzed. Key Words: cancer vaccines, antigens, immunity, resistance, therapy.  îáçîðå ðàññìîòðåíû ñîâðåìåííûå äàííûå î ðàçðàáîòêå âàêöèí ïðîòèâ çëîêà÷åñòâåííûõ íîâîîáðàçîâàíèé. Ïðåäñòàâëåíû ñâåäåíèÿ î çíà÷åíèè îïóõîëåñïåöèôè÷åñêèõ è îïóõîëåàññîöèèðîâàííûõ àíòèãåíîâ, ðîëè áåëêîâ ãëàâíîãî êîìïëåêñà ãèñòîñîâìåñòèìîñòè, ìîëåêóë àäãåçèè è ìîëåêóë, óñèëèâàþùèõ èììóííûé îòâåò â ôîðìèðîâàíèè ìåõàíèçìîâ óõîäà îïóõîëè èç-ïîä èììóííîãî íàäçîðà. Ïðîàíàëèçèðîâàíû ðåçóëüòàòû èññëåäîâàíèé ïî ñîçäàíèþ ðåêîìáèíàíòíûõ âàêöèí, à òàêæå âàêöèí íà îñíîâå íóêëåèíîâûõ êèñëîò è ãëèêîïåïòèäîâ. Ïðèâîäÿòñÿ äàííûå î ïåðñïåêòèâíîñòè èñïîëüçîâàíèÿ íîâûõ âåêòîðîâ äëÿ ââåäåíèÿ ãåíåòè÷åñêîãî ìàòåðèàëà â ãåíîì îïóõîëåâûõ êëåòîê, èñïîëüçîâàíèÿ ëèïîñîì è ôàêòîðîâ, óñèëèâàþùèõ èììóííûé îòâåò. Àíàëèçèðóþòñÿ êîíêðåòíûå ðåçóëüòàòû èñïîëüçîâàíèÿ íîâûõ ïðîòèâîîïóõîëåâûõ âàêöèí è ïóòè èõ äàëüíåéøåãî óñîâåðøåíñòâîâàíèÿ. Êëþ÷åâûå ñëîâà: ïðîòèâîîïóõîëåâûå âàêöèíû, àíòèãåíû, èììóíèòåò, ðåçèñòåíòíîñòü, òåðàïèÿ. Ïåðâûå ïîïûòêè îñóùåñòâèòü èììóíèçàöèþ ïðîòèâ çëîêà÷åñòâåííûõ îïóõîëåé áûëè ïðåäïðèíÿòû áîëåå 100 ëåò íàçàä [1, 2].  òî âðåìÿ óæå áûë äîñòèãíóò ðÿä ñåðüåçíûõ óñïåõîâ â áîðüáå ñ èíôåêöèîííûìè çàáîëåâàíèÿìè ìåòîäîì àêòèâíîé âàêöèíàöèè. Îäíàêî â îòíîøåíèè îïóõîëåé â áîëüøèíñòâå ñëó÷àåâ òàêèå ïîïûòêè îêàçàëèñü íåóäà÷íûìè. Òåðìèí ðàêîâûå âàêöèíû áûë ââåäåí èññëåäîâàòåëÿìè, èùóùèìè ïóòè èñïîëüçîâàíèÿ âîçìîæíîñòåé èììóííîé ñèñòåìû ïðè ëå÷åíèè áîëüíûõ îíêîëîãè÷åñêîãî ïðîôèëÿ è äëÿ ïðåäóïðåæäåíèÿ âîçíèêíîâåíèÿ çëîêà÷åñòâåííûõ íîâîîáðàçîâàíèé. Received: October 11, 1999. *Correspondence. E-mail: [email protected] Èñïîëüçóåìûå ñîêðàùåíèÿ: ÀÈÀÒ àíòèèäèîòèïè÷åñêèå àíòèòåëà; ÀÐÑ àíòèãåíïðåçåíòèðóþùèå êëåòêè; CD êëàñòåðû äèôôåðåíöèðîâêè; CEA êàðöèíî-ýìáðèîíàëüíûé àíòèãåí; CTL öèòîëèòè÷åñêèå Ò-ëèìôîöèòû; EGF-R ðåöåïòîð ýïèäåðìàëüíîãî ôàêòîðà ðîñòà; G-CSF ãðàíóëîöèòàðíî-ìàêðîôàãàëüíûé êîëîíèåñòèìóëèðóþùèé ôàêòîð; HLA ÷åëîâå÷åñêèé ëåéêîöèòàðíûé àíòèãåí; HSP áåëêè òåïëîâîãî øîêà; ÌÍÑ ãëàâíûé êîìïëåêñ ãèñòîñîâìåñòèìîñòè; NK-êëåòêè åñòåñòâåííûå êèëëåðíûå êëåòêè; ÒÀÀ îïóõîëåàññîöèèðîâàííûå àíòèãåíû; TCR ðåöåïòîð Ò-ëèìôîöèòà; TGF òðàíñôîðìèðóþùèé ôàêòîð ðîñòà; TSA îïóõîëåñïåöèôè÷åñêèå àíòèãåíû.  íàñòîÿùåå âðåìÿ áîëüøèíñòâî ïîäõîäîâ ê ñîçäàíèþ ïðîòèâîîïóõîëåâûõ âàêöèí íàïðàâëåíî íà äîñòèæåíèå ëå÷åáíîãî ýôôåêòà èëè ïðåäîòâðàùåíèå ðåöèäèâîâ ïîñëå õèðóðãè÷åñêîãî óäàëåíèÿ íîâîîáðàçîâàíèÿ èëè õèìèîëó÷åâîãî âîçäåéñòâèÿ. Ñïåöèôè÷åñêàÿ èììóíîòåðàïèÿ ïðåäïîëàãàåò èììóíèçàöèþ îïóõîëåâûìè êëåòêàìè ëèáî èçâëå÷åííûìè èç íèõ îïóõîëåâûìè àíòèãåíàìè (àêòèâíàÿ èììóíîòåðàïèÿ) èëè òðàíñïëàíòàöèþ èììóííûõ ëèìôîöèòîâ ëèáî ïîëó÷åííîé èç íèõ ÐÍÊ (àäîïòèâíàÿ èììóíîòåðàïèÿ). Âûäåëÿþò òðè ïðèíöèïèàëüíûõ êîìïîíåíòà, íåîáõîäèìûõ äëÿ ðåàëèçàöèè Ò-êëåòî÷íîãî èììóíèòåòà: 1) àíòèãåí, êîòîðûé ìîæåò áûòü ðàñïîçíàí ðåöåïòîðîì Ò-ëèìôîöèòà (TCR); 2) ýêñïðåññèÿ ãëàâíîãî êîìïëåêñà ãèñòîñîâìåñòèìîñòè (ÌÍÑ) â ñî÷åòàíèè ñ àíòèãåíîì; 3) êî-ñòèìóëèðóþùèå ñèãíàëû, èëè ïîìîùü, ïðåäñòàâëåííàÿ ýêçîãåííûìè èñòî÷íèêàìè ôàêòîðîâ ðîñòà. Ìåòîäû ìîëåêóëÿðíîé áèîëîãèè, âèðóñîëîãèè è èììóíîëîãèè ïîçâîëÿþò âûÿâèòü ðÿä àíòèãåíîâ â îïóõîëåâûõ êëåòêàõ ïðîäóêòû îíêîâèðóñîâ, ïðîäóêòû îíêîãåíîâ, ìóòàíòíûå áåëêè, áåëêè ýêòîïè÷åñêîãî ñèíòåçà, êëàñòåðû äèôôåðåíöèðîâêè (CD), áåëêè MHC ² è ²² êëàññîâ; áåëêè ïðîäóêòû â íîðìå ìîë÷àùèõ ãåíîâ. 100 Ýêñïåðèìåíòàëüíàÿ îíêîëîãèÿ 22, 99-109, 2000 (ñåíòÿáðü) ÌÅÕÀÍÈÇÌÛ ÓÕÎÄÀ ÎÏÓÕÎËÅÂÛÕ ÊËÅÒÎÊ ÈÇ-ÏÎÄ ÈÌÌÓÍÍÎÃÎ ÍÀÄÇÎÐÀ Îïóõîëü ìîæåò óõîäèòü èç-ïîä èììóííîãî íàäçîðà â ðåçóëüòàòå äåôåêòîâ â ëþáîì èç ïåðå÷èñëåííûõ âûøå çâåíüåâ, íåîáõîäèìûõ äëÿ ðàçâèòèÿ èììóííûõ ðåàêöèé. Ýòî ìîæåò ïðîèñõîäèòü ïðè áëîêàäå àíòèãåííûõ äåòåðìèíàíò îïóõîëåâûõ êëåòîê àíòèòåëàìè, ëèáî ðåöåïòîðîâ ëèìôîöèòîâ ðàñòâîðèìûìè àíòèãåíàìè, à òàêæå êîìïëåêñàìè àíòèãåí àíòèòåëî. Ïîêàçàíî òàêæå, ÷òî ïðè íèçêîì ñîîòíîøåíèè àíòèòåë è ëèìôîöèòîâ, ñ îäíîé ñòîðîíû, è îòíîñèòåëüíî áîëüøîãî ÷èñëà îïóõîëåâûõ êëåòîê ñ äðóãîé, ìîæåò íàáëþäàòüñÿ ñòèìóëÿöèÿ ðîñòà îïóõîëè. Îïóõîëåâûå àíòèãåíû íà ïîâåðõíîñòè îïóõîëåâûõ êëåòîê ìîãóò áûòü ìàñêèðîâàíû èììóíîëîãè÷åñêè èíåðòíûìè âåùåñòâàìè òèïà ìóêîïîëèñàõàðèäîâ, ãëèêîïåïòèäîâ è ò.ï.  ñëó÷àÿõ, åñëè îïóõîëåâûé ðîñò èíäóöèðóåòñÿ îíêîðíàâèðóñàìè, ïåðåäàþùèìèñÿ îò ðîäèòåëåé ïîòîìñòâó âåðòèêàëüíûì ïóòåì ÷åðåç ïîëîâûå êëåòêè, èëè ñ ìîëîêîì ìàòåðè, ëèáî òðàíñïëàöåíòàðíûì ïóòåì, ìîæåò âîçíèêíóòü òîëåðàíòíîñòü ê ïðîäóöèðóåìûì ýòèìè âèðóñàìè îïóõîëåâûì àíòèãåíàì.  îðãàíèçìå ñ ðàñòóùåé îïóõîëüþ îòìå÷àåòñÿ óâåëè÷åíèå êîëè÷åñòâà êëåòîê-ñóïðåññîðîâ.  îïûòàõ íà ìûøàõ ïîêàçàíî, ÷òî ýòè êëåòêè èíãèáèðóþò èììóííûé îòâåò ñèëüíåå, ÷åì ñóïðåññîðû, ïîëó÷åííûå îò çäîðîâûõ æèâîòíûõ. Ðàñòóùàÿ îïóõîëü ïðîäóöèðóåò èììóíîäåïðåññèâíûå ôàêòîðû, áëîêèðóþùèå ôóíêöèè Ò-ëèìôîöèòîâ, íàòóðàëüíûõ êèëëåðíûõ êëåòîê è ìàêðîôàãîâ. Ïðè ïðîâåäåíèè õèìèîòåðàïèè ïðîòèâîîïóõîëåâûìè ïðåïàðàòàìè îòìå÷àåòñÿ òàêæå çíà÷èòåëüíîå ïîäàâëåíèå èììóííûõ ðåàêöèé. Âûÿñíåíèå ìîëåêóëÿðíûõ ìåõàíèçìîâ, âåäóùèõ ê óõîäó îïóõîëè èç-ïîä èììóííîãî íàäçîðà, ìîæåò îêàçàòüñÿ ïîëåçíûì â ðàçðàáîòêå ìåòîäîâ ïðîòèâîðàêîâîé òåðàïèè. ×àñòî íàáëþäàåìîå ïîäàâëåíèå ýêñïðåññèè ìîëåêóë ÌÍÑ â êëåòêàõ îïóõîëåé ðàçíîé ïðèðîäû [3, 4] ÿâëÿåòñÿ ãëàâíîé ïðè÷èíîé, îãðàíè÷èâàþùåé óñïåøíóþ èììóíîòåðàïèþ, îñíîâàííóþ íà äåéñòâèè öèòîëèòè÷åñêèõ Ò-ëèìôîöèòîâ (ÑÒL) [5, 6]. Ìåõàíèçì óõîäà èç-ïîä èììóííîãî íàäçîðà â ðÿäå ñëó÷àåâ ñâÿçàí ñ íàëè÷èåì â îïóõîëè îïóõîëåàññîöèèðîâàííûõ àíòèãåíîâ (ÒÀÀ).  ñâÿçè ñ òåì, ÷òî íîðìàëüíûå òêàíè òàêæå ïðîäóöèðóþò ýòè àíòèãåíû, âîçíèêàåò òîëåðàíòíîñòü ê êëåòêàì, íåñóùèì ýòè àíòèãåíû, â òîì ÷èñëå è ê îïóõîëåâûì, ñâÿçàííàÿ ëèáî ñ îòñóòñòâèåì, ëèáî ñ àíåðãèåé ñîîòâåòñòâóþùèõ ñóáïîïóëÿöèé Ò-ëèìôîöèòîâ [7, 8].  òî æå âðåìÿ àðåàêòèâíîñòü ïî îòíîøåíèþ ê ÒÀÀ íå ÿâëÿåòñÿ àáñîëþòíîé ó ìíîãèõ áîëüíûõ îíêîëîãè÷åñêîãî ïðîôèëÿ è ó çäîðîâûõ ëèö ñóùåñòâóåò èììóííûé îòâåò íà ýòè àíòèãåíû, â ñâÿçè ñ ÷åì èìååòñÿ âîçìîæíîñòü âàêöèíàöèè ñ ïîìîùüþ òàêèõ àíòèãåíîâ. Ïðè ýòîì íå èñêëþ÷àåòñÿ, ÷òî èììóííûå ðåàêöèè ïðîòèâ îïàñíûõ ñîáñòâåííûõ àíòèãåíîâ, èìåþùèõñÿ è â íîðìå, ïðè âàêöèíàöèè ïîäàâëÿþòñÿ [9]. Êðîìå òîãî, ïëîòíîñòü ÒÀÀ íà ïîâåðõíîñòè íîðìàëüíûõ êëåòîê íåáîëüøàÿ, â ñâÿçè ñ ÷åì îíè íå ïîâðåæäàþòñÿ àíòèòåëàìè èëè Ò-êëåòêàìè, è äîñòàòî÷íî âûñîêàÿ íà îïóõîëåâûõ êëåòêàõ, èç-çà ÷åãî îíè ðàçðóøàþòñÿ ýòèìè ýôôåêòîðàìè [10]. Ê òîìó æå èíôèëüòðàöèÿ ìàêðîôàãàìè íîðìàëüíûõ òêàíåé çíà÷èòåëüíî íèæå, ÷åì îïóõîëåâûõ. Ê ÒÀÀ, âûÿâëåííûì â êàðöèíîìàõ, îòíîñÿòñÿ: CO171A/ GA733/ KSA/EpCAM [11], p53 [12, 13], MUC-1 [1416], ê àññîöèèðîâàííûì ñ ìåëàíîìàìè gp-100 [17], MART-1/ MELANA [18]. Îïóõîëåñïåöèôè÷åñêèå àíòèãåíû (TSA), ïðîäóöèðóåìûå òîëüêî â îïóõîëåâûõ êëåòêàõ, ïðåäñòàâëÿþò ñîáîé ëèáî ìóòèðîâàííûå èñõîäíûå áåëêè, íàïðèìåð, mð53[19], mras [20], m-β-êàòåíèí [21], mEGF-R, ëèáî àíòèãåíû Å6 è Å7, èíäóöèðîâàííûå âèðóñàìè, íàïðèìåð, âèðóñîì ïàïèëëîìàòîçà [22]. Îòñóòñòâèå ýòèõ áåëêîâ â íîðìàëüíûõ òêàíÿõ îáåñïå÷èâàåò è îòñóòñòâèå òîëåðàíòíîñòè ê íèì, ÷òî ìîæåò áûòü èñïîëüçîâàíî â ìåõàíèçìàõ èììóííîé çàùèòû.  ýêñïåðèìåíòàõ íà æèâîòíûõ ïîêàçàíî, ÷òî ïåïòèäû m-ras è m-EGFR ìîãóò áûòü ïðèìåíåíû â êà÷åñòâå ïðîòèâîîïóõîëåâûõ âàêöèí [23, 24].  òî æå âðåìÿ âûÿñíèëîñü, ÷òî TSA, ïðîäóöèðóåìûå ðàñòóùåé îïóõîëüþ, â ðÿäå ñëó÷àåâ îêàçûâàþòñÿ íåèììóíîãåííûìè [25].  ýòèõ èññëåäîâàíèÿõ ïîêàçàíî, ÷òî îïóõîëåâûå êëåòêè ìûøåé, ýêñïðåññèðîâàâøèå ýïèòîï TSA, êîäèðóåìûé âèðóñîì ëèìôîöèòàðíîãî õîðèîìåíèíãèòà, àêòèâèðóþò Ò-ëèìôîöèòû in vitro, îäíàêî íå èíäóöèðóþò èõ ïðîòèâîîïóõîëåâóþ àêòèâíîñòü in vivo.  òî æå âðåìÿ, åñëè èììóíèçàöèþ ïðîâîäèëè àíòèãåíîì, ñâÿçàííûì ñ äåíäðèòíûìè êëåòêàìè, íàñòóïàëà áûñòðàÿ ðåãðåññèÿ îïóõîëè. Óõîä îïóõîëè èç-ïîä èììóííîãî êîíòðîëÿ ïðîèñõîäèò ïðè ðàçëè÷íûõ óñëîâèÿõ. Âî-ïåðâûõ, â ïðîöåññå îïóõîëåâîãî ðîñòà èëè èììóíîòåðàïèè âîçìîæíà ïîòåðÿ ñîîòâåòñòâóþùèõ àíòèãåíîâ (òàê íàçûâàåìûé âàðèàíò ïîòåðè àíòèãåíà). Âî-âòîðûõ, ìîãóò âîçíèêàòü ðàçëè÷íûå äåôåêòû íà ïóòÿõ ïðîöåññèíãà àíòèãåíà â ðåçóëüòàòå èíòåðôåðåíòíûõ âçàèìîîòíîøåíèé ïðè îáðàçîâàíèè èììóíîãåííûõ ïåïòèäîâ â öèòîçîëå àíòèãåíïðåçåíòèðóþùèõ êëåòîê (ÀÐÑ) (íàïðèìåð, ïðè äåôåêòàõ îáðàçîâàíèÿ ïðîòåàñîì) ëèáî â ðåçóëüòàòå íåàäåêâàòíîãî ââåäåíèÿ ïåïòèäîâ â ýíäîïëàçìàòè÷åñêèé ðåòèêóëóì (íàïðèìåð, ïðè äåôåêòàõ â ïåðåíîñ÷èêå, ñâÿçàííîì ñ ïðîöåññèíãîì àíòèãåíà). Â-òðåòüèõ, ïîäàâëåíèå èëè îòñóòñòâèå ñîîòâåòñòâóþùåé àëëåëè HLA ìîæåò ïðåäîòâðàòèòü ïðåçåíòàöèþ ïåïòèäà â ñîåäèíåíèè ñ ÌÍÑ íà êëåòî÷íîé ïîâåðõíîñòè. Êðîìå òîãî, â íåêîòîðûõ ñëó÷àÿõ àðåàêòèâíîñòè îáíàðóæèâàþò ìóòàöèè â ñòðóêòóðå ãåíà β2-ìèêðîãëîáóëèíà. Åùå îäèí ïóòü óõîäà èç-ïîä èììóííîãî êîíòðîëÿ ñâÿçàí ñ èíäóêöèåé àïîïòîçà Fas+ β CTL â ðåçóëüòàòå ýêñïðåññèè ñîîòâåòñòâóþùèõ ëèãàíäîâ íà îïóõîëåâûõ êëåòêàõ, íàïðèìåð Fas L [26, 27]. Òàê æå ìîãóò óíè÷òîæàòüñÿ è äåíäðèòíûå êëåòêè. Íàêîíåö, îïóõîëåâûå êëåòêè ìîãóò èíäóöèðîâàòü àïîïòîç â àêòèâèðîâàííûõ Ò-ëèìôîöèòàõ çà ñ÷åò ñáðàñûâàíèÿ ìîëåêóë HLA ñî ñâîåé ïîâåðõíîñòè. Experimental Oncology 22, 99-109, 2000 (September) Îäíèì èç ìåõàíèçìîâ óõîäà îïóõîëè èç-ïîä èììóííîãî êîíòðîëÿ ìîæåò áûòü ñíèæåíèå ýêñïðåññèè èëè äåôåêò ïðîäóêöèè ôàêòîðîâ àäãåçèè [28]. Ñðåäè ïîñëåäíèõ îñîáîå çíà÷åíèå èìååò ñóïåðñåìåéñòâî èíòåãðèíîâ è ñîîòâåòñòâóþùèå èì ëèãàíäû. Èíòåãðèíû ñåìåéñòâî ãåòåðîäèìåðîâ, ñîäåðæàùèõ îáùóþ β-öåïü (CD18) è îäíó èç òðåõ ðàçëè÷íûõ α-öåïåé (CD11a, CD11b, CD11c). Êðîìå òîãî, îïèñàíû äðóãèå èíòåãðèíû, âûÿâëÿåìûå àíòèòåëàìè ïðîòèâ CD2 è CD56 [28]. Ïîêàçàíî, ÷òî áëîêèðîâàíèå ìîëåêóë àäãåçèè èëè àäåêâàòíûõ èì ëèãàíäîâ ñîîòâåòñòâóþùèìè ìîíîêëîíàëüíûìè àíòèòåëàìè ÷àñòè÷íî èëè ïîëíîñòüþ ïðåäîòâðàùàåò ëèçèñ îïóõîëåâûõ êëåòîê CTL èëè NK-êëåòêàìè [29, 30]. Âàæíóþ ðîëü â ðåàëèçàöèè ýôôåêòîðíûõ ôóíêöèé ëèìôîöèòîâ èãðàþò ìåìáðàííûå ñòðóêòóðû, àêòèâèðóþùèå ëèçèñ êëåòîê-ìèøåíåé. Ñðåäè íèõ îñîáî âàæíûìè ÿâëÿþòñÿ ìîëåêóëû CD16 [31], NKG2 [32,33], CD69 [34], gp42 [35], 2B4 [36], p38 [37]. Áëîêèðîâàíèå ýòèõ ìîëåêóë òàêæå ïðåäîòâðàùàåò ëèçèñ îïóõîëåâûõ êëåòîê ìîëåêóëàìè-ýôôåêòîðàìè [38]. Ôàêòîðû, îñëàáëÿþùèå ìåæêëåòî÷íûå âçàèìîäåéñòâèÿ, ðåçêî ñíèæàþò ýôôåêòèâíîñòü èììóííûõ ðåàêöèé. Íàïðèìåð, âûñîêàÿ êîíöåíòðàöèÿ ïîëèàìèíîâ íà ïîâåðõíîñòè îïóõîëåâûõ êëåòîê ñïîñîáíà ýêðàíèðîâàòü îòðèöàòåëüíî çàðÿæåííûå äåòåðìèíàíòû, â òî âðåìÿ êàê ïîëèñàõàðèäíûå èëè îëèãîíóêëåîòèäíûå ôðàãìåíòû ìîëåêóë íà êëåòî÷íûõ ìåìáðàíàõ è â ìåæêëåòî÷íîì ïðîñòðàíñòâå áëîêèðóþò ïîëîæèòåëüíî çàðÿæåííûå ôðàãìåíòû àíòèãåííûõ äåòåðìèíàíò. Ðàñòóùàÿ îïóõîëü ïðîäóöèðóåò èììóíîñóïðåññèâíûå ôàêòîðû, ê êîòîðûì îòíîñÿòñÿ TGF-β, èíòåðëåéêèí-10 è äðóãèå, êîòîðûå ìîãóò ïîäàâëÿòü àêòèâíîñòü Ò-ëèìôîöèòîâ [39, 40]. Êðîìå òîãî, îïóõîëåâûå êëåòêè ìîãóò ýêñïðåññèðîâàòü Fas-ëèãàíä, èíäóöèðóþùèé àïîïòîç Ò-ëèìôîöèòîâ [41]. ×òîáû ïðåîäîëåòü èíãèáèðóþùèé ýôôåêò ôàêòîðîâ ðîñòà è äðóãèõ öèòîêèíîâ, ðàçðàáîòàíà ñòðàòåãèÿ èììóíèçàöèè îïóõîëåâûìè êëåòêàìè, òðàíñäóöèðîâàííûìè àíòèñåíñîâîé TGF-β-êîìïëåìåíòàðíîé (ñ) ÄÍÊ [42], ëèáî (ñ) ÄÍÊ, êîäèðóþùåé èììóíîñòèìóëèðóþùèå öèòîêèíû [43]. Ýòîò ïîäõîä îêàçàëñÿ ýôôåêòèâíûì íåñìîòðÿ íà òî, ÷òî îïóõîëåâûå êëåòêè ïðîäîëæàëè ïðîäóöèðîâàòü öèòîêèíû è ôàêòîðû ðîñòà. Êî-ñòèìóëèðóþùèå ìîëåêóëû, òàêèå, êàê Â71 è Â72, ýêñïðåññèðóåìûå îïóõîëåâûìè êëåòêàìè èëè ÀÐÑ, ñïîñîáñòâóþò èíäóêöèè ýôôåêòèâíîãî ïðîòèâîîïóõîëåâîãî Ò-êëåòî÷íîãî îòâåòà [44].  ïðîòèâîïîëîæíîñòü íîðìàëüíûì êëåòêàì îïóõîëåâûå êëåòêè ìåíåå àêòèâíî ïðîäóöèðóþò ôàêòîð Â7. Îáíàðóæåíèå íîâûõ ôàêòîðîâ èììóíîäåïðåññèè èëè ìåõàíèçìîâ óõîäà îïóõîëåâûõ êëåòîê èç-ïîä èììóííîãî íàäçîðà ñòèìóëèðîâàëî ïîïûòêè ðàçðàáîòàòü íîâûå ïðîòèâîîïóõîëåâûå âàêöèíû. Òàê, áûëî óñòàíîâëåíî, ÷òî âàêöèíàöèÿ îïóõîëåâûìè êëåòêàìè, òðàíñäóöèðîâàííûìè ôàêòîðîì Â7, èíäóöèðóåò ïðîòèâîîïóõîëåâóþ çàùèòó [4549].  ýòèõ ðàáîòàõ ïîêàçàíî, ÷òî Â7 íåîáõîäèì äëÿ àêòèâàöèè Ò-ëèìôîöèòîâ, íî íå äëÿ èõ ýôôåêòîðíûõ ôóíê- 101 öèé, â ñâÿçè ñ ÷åì îíè îêàçûâàþòñÿ ýôôåêòèâíûìè òàêæå è ïðîòèâ îïóõîëåâûõ êëåòîê, ìàëî ïðîäóöèðóþùèõ èëè âîîáùå íå ïðîäóöèðóþùèõ ôàêòîð Â7. ÂÀÊÖÈÍÛ ÍÀ ÎÑÍÎÂÅ ÖÅËÜÍÛÕ ÊËÅÒÎÊ È ÊËÅÒÎ×ÍÛÕ ËÈÇÀÒΠÏåðâûìè ïîïûòêàìè ïðîòèâîîïóõîëåâîé âàêöèíàöèè áûëè ðàáîòû ñ èñïîëüçîâàíèåì öåëüíûõ îïóõîëåâûõ êëåòîê, îäíàêî ââåäåíèå â îðãàíèçì æèâûõ íåïîâðåæäåííûõ êëåòîê íåäîïóñòèìî èç-çà îïàñíîñòè ðàñïðîñòðàíåíèÿ ïðîöåññà. Èñïûòàâàëè ïðîòèâîîïóõîëåâóþ âàêöèíó, ïðèãîòîâëåííóþ èç öåëüíûõ êëåòîê, ó áîëüíûõ ñ êîëîðåêòàëüíûì ðàêîì [50]. Ïðè ýòîì áûëè èñïîëüçîâàíû àóòîëîãè÷íûå îïóõîëåâûå êëåòêè, óáèòûå îáëó÷åíèåì, â ñî÷åòàíèè ñ âàêöèíîé BCG. Äëÿ êîíòðîëÿ äðóãèì áîëüíûì ââîäèëè îáëó÷åííûå îïóõîëåâûå êëåòêè áåç BCG.  òå÷åíèå òðåõ ëåò 2 áîëüíûõ èç 6 áûëè êëèíè÷åñêè çäîðîâû, ó îñòàëüíûõ ðàçâèëèñü ðåöèäèâû ðàêà. Ïîëîæèòåëüíûé ýôôåêò ïîëó÷åí ïðè èñïîëüçîâàíèè âàêöèíû èç öåëüíûõ îïóõîëåâûõ êëåòîê â ýêñïåðèìåíòàõ íà æèâîòíûõ ñ îïóõîëÿìè ìîëî÷íîé æåëåçû [51]. Îáíàäåæèâàþùèå ðåçóëüòàòû ïîëó÷åíû ïðè âàêöèíàöèè öåëüíûìè êëåòêàìè áîëüíûõ c ìåëàíîìîé [52]. Îòêðûòèå àññîöèèðîâàííûõ ñ ìåëàíîìîé àíòèãåíîâ âîçîáíîâèëî èíòåðåñ ê ïåïòèäíûì àíòèãåíàì îïóõîëåé è ïåïòèäàì, íàãðóæåííûì íà ÀÐÑ, êîòîðûå ñòàëè îñíîâîé ðàçðàáîòêè âàêöèí íîâîãî ïîêîëåíèÿ. Ëèçàòû àóòîëîãè÷íûõ îïóõîëåâûõ êëåòîê, ïîëó÷åííûõ îò áîëüíûõ ñ ìåëàíîìîé, èñïîëüçîâàëè äëÿ âíóòðèêîæíîé èììóíèçàöèè [53]. Ñåìíàäöàòè áîëüíûì c ìåëàíîìîé ââîäèëè ïî âîçðàñòàþùåé øêàëå 105107 àóòîëîãè÷íûõ ÀÐÑ 1 ðàç â ìåñÿö íà ïðîòÿæåíèè 4 ìåñ. Ó îäíîãî áîëüíîãî íàáëþäàëè ÷àñòè÷íîå ðàññàñûâàíèå ïîäêîæíîãî óçëà, 13 áîëüíûõ æèëè îò 4 äî 30 ìåñ (â ñðåäíåì 17 ìåñ) ïîñëå îêîí÷àíèÿ êóðñà èììóíèçàöèè. Ðàíåå áûëà ïîêàçàíà âîçìîæíîñòü ïîëó÷åíèÿ ïîëèâàëåíòíîé âàêöèíû èç ñìûâîâ êóëüòèâèðóåìûõ ìåëàíîìíûõ êëåòîê ÷åëîâåêà [54]. Òðè îáðàçöà ïðèãîòîâëåííîé òàêèì îáðàçîì âàêöèíû èìåëè îäèíàêîâûé ñïåêòð àíòèãåíîâ. Âàêöèíà áûëà èñïûòàíà íà 13 áîëüíûõ ïðè âíóòðèêîæíîé èììóíèçàöèè àíòèãåíîì â âîçðàñòàþùèõ äîçàõ, ãóìîðàëüíûé îòâåò îòìå÷åí ó 5 (38%), êëåòî÷íûé ó 4 (31%). Ó îäíîãî áîëüíîãî ïîëíîñòüþ ðàññîñàëèñü êîæíûå ìåòàñòàçû, ó äðóãîãî îíè îñòàâàëèñü áåç èçìåíåíèé â òå÷åíèå 14 ìåñ. Ïðîãðåññ â èçó÷åíèè îïóõîëåâûõ àíòèãåíîâ è âûÿñíåíèå ìîëåêóëÿðíûõ ìåõàíèçìîâ âçàèìîäåéñòâèÿ îïóõîëè è îðãàíèçìà ñòèìóëèðîâàë è èññëåäîâàíèÿ ïî ðàçðàáîòêå ïðîòèâîîïóõîëåâûõ âàêöèí íîâîãî ïîêîëåíèÿ. Èñïîëüçîâàíèå òåõíîëîãèè ïåðåíîñà ãåíîâ ïðèâåëî ê ñîçäàíèþ âàêöèí íà îñíîâå êëåòîê, ýêñïðåññèðóþùèõ ïîâûøåííûå êîëè÷åñòâà íàòèâíûõ è ìîäèôèöèðîâàííûõ ôàêòîðîâ ðîñòà, ìîëåêóë àäãåçèè è êî-ñòèìóëèðóþùèõ ìîëåêóë [55]. Ñóïåðàíòèãåíû (Sag) ýòî ñåìåéñòâî áàêòåðèàëüíûõ è âèðóñíûõ áåëêîâ ñ ìîùíûìè èììóíîñòèìóëèðóþùèìè ñâîéñòâàìè, êîòîðûå ñâÿçûâàþòñÿ ñ ìîëåêóëàìè MHC ²² êëàññà è àêòèâèðóþò 102 Ýêñïåðèìåíòàëüíàÿ îíêîëîãèÿ 22, 99-109, 2000 (ñåíòÿáðü) Ò-ëèìôîöèòû [56]. Ñóïåðàíòèãåíû òàêæå èíäóöèðóþò ïðîäóêöèþ áîëüøîãî êîëè÷åñòâà öèòîêèíîâ è ïîâûøàþò öèòîòîêñè÷íîñòü CD4+- è CD8+-Ò-ëèìôîöèòîâ. Ñêîíñòðóèðîâàí êîìáèíèðîâàííûé áåëîê, ñîñòîÿùèé èç ìîíîêëîíàëüíûõ àíòèòåë ïðîòèâ îïóõîëåâîãî àíòèãåíà (C215 Fab) è áàêòåðèàëüíîãî ýíòåðîòîêñèíà À (SEA) [56].  îïûòàõ íà ìûøàõ áûë ïîêàçàí âûðàæåííûé ïðîòèâîîïóõîëåâûé ýôôåêò ïðè èììóíèçàöèè ýòèì áåëêîì. Ðÿä äàííûõ ñâèäåòåëüñòâóåò î òîì, ÷òî èììóíîãåííîñòü ïîëèïåïòèäîâ çíà÷èòåëüíî âîçðàñòàåò, åñëè ê ïîëèïåïòèäíîé öåïè ïðèñîåäèíåí óãëåâîäíûé êîìïîíåíò.  ñâÿçè ñ ýòèì áûëè ñîçäàíû èñêóññòâåííûå êîíúþãàòû ïîëèïåïòèäîâ ñ óãëåâîäíûì îñòàòêîì. Ðàçðàáîòàí ïðîñòîé è ýôôåêòèâíûé ìåòîä ïðèñîåäèíåíèÿ áåëêîâîãî ïðîäóêòà STn (Neu Ac2), ñóïåðýêñïðåññèðóåìîãî êëåòêàìè ðÿäà îïóõîëåé ÷åëîâåêà, ê ãåìîöèàíèíó ïðóäîâèêà (KLH) [57]. Òàêîé ïðîäóêò îáåñïå÷èâàåò âûñîêèé óðîâåíü ñèíòåçà ñïåöèôè÷åñêèõ àíòèòåë ïðîòèâ STn è ìîæåò èñïîëüçîâàòüñÿ â ñõåìàõ ïðîòèâîîïóõîëåâîé âàêöèíîòåðàïèè. Ðåêîìáèíàíòíûé áåëîê Å7 âèðóñà ïàïèëëîìàòîçà ÷åëîâåêà HPV16 â ñî÷åòàíèè ñ ñèíòåòè÷åñêèì àäúþâàíòîì PROVAX áûë èñïûòàí â êà÷åñòâå âàêöèíû íà ìûøàõ ñ ìåòàñòàçèðóþùåé ìåëàíîìîé HOPE 2 [58]. Ïîêàçàíà çíà÷èòåëüíàÿ çàäåðæêà ðîñòà îïóõîëè, ïðè ýòîì îñíîâíûìè ýôôåêòîðíûìè êëåòêàìè áûëè ÑD8+-Ò-ëèìôîöèòû. D. Berd et al. [59] ðàçðàáîòàëè ñïîñîá âàêöèíîòåðàïèè ñ èñïîëüçîâàíèåì îïóõîëåâûõ êëåòîê ñ ïðèñîåäèíåííûì ñèíòåòè÷åñêèì ãàïòåíîì äèíèòðîôåíîëîì. Ìîäèôèöèðîâàííûå ýòèì ãàïòåíîì îïóõîëåâûå êëåòêè â ñìåñè ñ âàêöèíîé BCG ìíîãîêðàòíî ââîäèëè âíóòðèêîæíî áîëüíûì ñ ìåòàñòàçèðóþùåé ìåëàíîìîé, ÷òî âûçûâàëî âîñïàëåíèå â îïóõîëåâûõ óçëàõ. Ïðè ãèñòîëîãè÷åñêîì èññëåäîâàíèè â ýòèõ óçëàõ îòìå÷àëè èíôèëüòðàöèþ Ò-ëèìôîöèòàìè, áîëüøèíñòâî èç êîòîðûõ áûëè ÑD8+. Ýòè êëåòêè àêòèâíî ïðîäóöèðîâàëè ãàììà-èíòåðôåðîí in situ, â íèõ îáíàðóæåíà óíèêàëüíàÿ íîâàÿ ñòðóêòóðà TCR. Ïðè ýòîì íàáëþäàëè ðåãðåññèþ ìåòàñòàçîâ, íàèáîëåå âûðàæåííóþ â ëåãêèõ. Èç 62 áîëüíûõ c ìåëàíîìîé ²²² ñòàäèè ïîñëå ïðåäâàðèòåëüíîé ëèìôàäåíýêòîìèè áîëåå 5 ëåò ïðîæèëè 45%, à ïîêàçàòåëü îáùåé ïðîäîëæèòåëüíîñòè æèçíè óâåëè÷èëñÿ íà 58%. Ñõîäíûå ðåçóëüòàòû áûëè ïîëó÷åíû ïðè òàêîì ìåòîäå ëå÷åíèÿ áîëüíûõ ðàêîì ÿè÷íèêîâ ²²² ñòàäèè. Èñïîëüçóÿ ìîäåëü ìàñòîöèòîìû Ð815 è ìåëàíîìû Ì3, W. Schmidt et al. [60] èñïûòàëè â ëå÷åáíûõ öåëÿõ íà ìûøàõ ÷åòûðå ðàçëè÷íûõ îïóõîëåâûõ àíòèãåíà îòäåëüíî è â ñî÷åòàíèè ñ àäúþâàíòîì Ôðåéíäà. Íè â îäíîì èç îïûòîâ íå áûëî ïîëó÷åíî ïîëîæèòåëüíîãî ðåçóëüòàòà, îäíàêî èñïîëüçîâàíèå ýòèõ àíòèãåíîâ â ñî÷åòàíèè ñ ïîëèêàòèîíîì ïîëèëèçèíîì äàëî ìîùíûé ïðîòèâîîïóõîëåâûé ýôôåêò, êîòîðûé çàâèñåë îò òèïà ïîëèëèçèíà. Àâòîðû îáúÿñíÿþò ïîëó÷åííûå ðåçóëüòàòû òåì, ÷òî ïîëîæèòåëüíî çàðÿæåííûå ìîëåêóëû ïîëèëèçèíà îáåñïå÷èâàþò èìïîðò êîðîòêèõ ïîëèïåïòèäíûõ ëèãàíäîâ âíóòðü ÀÐÑ. Èñõîäÿ èç äàííûõ î òîì, ÷òî êîðîâûå ïåïòèäû ìóöèíîâ, ïðîäóöèðóåìûõ îïóõîëåâûìè êëåòêàìè, ñóùåñòâåííî îòëè÷àþòñÿ îò ìóöèíîâ, ñåêðåòèðóåìûõ êëåòêàìè íîðìàëüíîãî ýïèòåëèÿ, êèòàéñêèå èññëåäîâàòåëè [61] âûäåëèëè êîðîâîé ïåïòèä (àïîìóöèí) èç êóëüòèâèðóåìûõ êëåòîê ðàêà ïîäæåëóäî÷íîé æåëåçû ìûøåé SW 1990.  ñî÷åòàíèè ñ àäúþâàíòîì âûäåëåííûé ïåïòèä, òàê æå, êàê è ñèíòåòè÷åñêèå ïåïòèäû MUC-2 è MUC-3, èíäóöèðîâàëè êëåòî÷íûé è ãóìîðàëüíûé îòâåò ó ìûøåé, ðåãèñòðèðóåìûé ïî ïîÿâëåíèþ àíòèòåë â ðåàêöèè ELISA, è êëåòî÷íûé èììóíèòåò, ðåãèñòðèðóåìûé â ðåàêöèè ãèïåð÷óâñòâèòåëüíîñòè çàìåäëåííîãî òèïà. Ñåëåçåíî÷íûå êëåòêè âàêöèíèðîâàííûõ ìûøåé ïðîÿâëÿëè öèòîòîêñè÷åñêóþ àêòèâíîñòü ïðîòèâ êëåòîê ðàêà ÷åëîâåêà, â òîì ÷èñëå ðàêà òîëñòîãî êèøå÷íèêà, æåëóäêà, ïîäæåëóäî÷íîé æåëåçû è ëåéêîçíûõ êëåòîê. J. Samuel et al. [62] äëÿ èììóíèçàöèè ìûøåé èñïîëüçîâàëè êëåòêè ìûøèíîé àäåíîêàðöèíîìû GZHI, â êîòîðûå áûë òðàíñôåöèðîâàí ÷åëîâå÷åñêèé àíòèãåí MUC 1, çàêëþ÷åííûé â ëèïîñîìû. Òàêàÿ èììóíèçàöèÿ îáåñïå÷èâàëà ïîëíóþ çàùèòó æèâîòíûõ êàê ïðè ïîäêîæíîé, òàê è ïðè âíóòðèâåííîé ïðèâèâêå îïóõîëåâûõ êëåòîê. Ïðè ýòîì ïðîëèôåðèðóþùèå Ò-ëèìôîöèòû ïðîäóöèðîâàëè çíà÷èòåëüíîå êîëè÷åñòâî ãàììà-èíòåðôåðîíà. ÂÀÊÖÈÍÛ ÍÀ ÎÑÍÎÂÅ ÀÍÒÈÈÄÈÎÒÈÏÈ×ÅÑÊÈÕ ÀÍÒÈÒÅË Îäíèì èç ïóòåé ñîçäàíèÿ ïðîòèâîîïóõîëåâûõ âàêöèí áûëî èñïîëüçîâàíèå äëÿ èõ ïðèãîòîâëåíèÿ ãàíãëèîçèäîâ èç ïëàçìàòè÷åñêèõ ìåìáðàí îïóõîëåâûõ êëåòîê [63]. Äðóãîé ïîäõîä áûë ñâÿçàí ñ èñïîëüçîâàíèåì ïðîòèâîäèñèàëîãàíãëèîçèäíûõ àíòèèäèîòèïè÷åñêèõ àíòèòåë (ÀÈÀÒ) [64]. ÀÈÀÒ ïðîòèâ äèñèàëîãàíãëèîçèäà GD2 ïîëíîñòüþ èìèòèðóþò äåéñòâèå èñõîäíîãî äèñèàëîãàíãëèîçèäà. Ýòî ñâÿçàíî ñ òåì, ÷òî ÀÈÀÒ âîñïðîèçâîäÿò êîíôîðìàöèþ èñõîäíîé àíòèãåííîé äåòåðìèíàíòû. Äâåíàäöàòè áîëüíûì ââîäèëè 1, 2, 4 èëè 8 ìã ÀÈÀÒ â ñìåñè ñ àäúþâàíòîì QS-21 (ðàç â íåäåëþ â òå÷åíèå ìåñÿöà, à çàòåì îäèí ðàç â ìåñÿö). Ó îäíîãî áîëüíîãî íàáëþäàëè ïîëíóþ êëèíè÷åñêóþ ðåìèññèþ, ó øåñòè ðåìèññèÿ äëèëàñü îò 9 äî 23 ìåñ.  äðóãîì èññëåäîâàíèè [65] áûëè ïîëó÷åíû ÀÈÀÒ ïðîòèâ êàðöèíî-ýìáðèîíàëüíîãî àíòèãåíà (ÑÅÀ). Ïîñëå áåçóñïåøíîé ñòàíäàðòíîé òåðàïèè 24 áîëüíûì ñ çàïóùåííûì ðàêîì òîëñòîãî êèøå÷íèêà ââîäèëè ïî 1, 2 èëè 4 ìã ÀÈÀÒ, ñîðáèðîâàííûõ íà ãèäðîîêèñè àëþìèíèÿ ðàç â íåäåëþ â òå÷åíèå ìåñÿöà, à çàòåì ðàç â ìåñÿö ïîêà óâåëè÷èâàëàñü îïóõîëü.  ñûâîðîòêå êðîâè ó 17 áîëüíûõ èç 23 áûëè íàéäåíû ÀÈÀÒ, à ó 13 èç íèõ èñòèííûå àíòè-ÑÅÀ-àíòèòåëà. Ïîñëåäíèå áûëè ïîëèêëîíàëüíûìè, à ó 11 áîëüíûõ íàáëþäàëàñü îïîñðåäîâàííàÿ àíòèòåëàìè êëåòî÷íàÿ öèòîòîêñè÷íîñòü. Ó 10 áîëüíûõ âûÿâëåíà èäèîòèïè÷åñêàÿ Ò-êëåòî÷íàÿ àêòèâíîñòü, à ó 5 ñïåöèôè÷åñêàÿ àêòèâíîñòü Experimental Oncology 22, 99-109, 2000 (September) ïðîòèâ ÑÅÀ. Íè ó îäíîãî áîëüíîãî íå áûëî êëèíè÷åñêîãî óëó÷øåíèÿ, îäíàêî ïðèìåíåíèå ÀÈÀÒ ïîçâîëèëî, ïî ìíåíèþ àâòîðîâ, âçëîìàòü èììóíîëîãè÷åñêóþ òîëåðàíòíîñòü ó áîëüøèíñòâà áîëüíûõ. Äëèòåëüíîñòü æèçíè áîëüíûõ, ëå÷åííûõ ÀÈÀÒ, îêàçàëàñü ñðàâíèìîé ñ äëèòåëüíîñòüþ æèçíè áîëüíûõ, ïîëó÷àâøèõ äðóãèå âèäû òåðàïèè, â òîì ÷èñëå ñ ïðèìåíåíèåì èðèíîòåêàíà, íî òÿæåñòü ïðîÿâëåíèÿ òîêñè÷íîñòè ïðè ýòîì áûëà çíà÷èòåëüíî íèæå.  êà÷åñòâå îáùåãî ÒÀÀ ìîæåò áûòü èñïîëüçîâàí ìóòàíòíûé áåëîê ð53. Îñîáåííî öåííûì â ýòîì íàïðàâëåíèè ìîæåò îêàçàòüñÿ èñïîëüçîâàíèå íåäàâíî ðàçðàáîòàííîãî ìåòîäà ïîëó÷åíèÿ áåëêà ð53 è àóòîàíòèòåë èç êðîâè ÷åëîâåêà â ïðåïàðàòèâíûõ êîëè÷åñòâàõ [66]. ÁÅËÊÈ ÒÅÏËÎÂÎÃÎ ØÎÊÀ Ïîñêîëüêó ñåìåéñòâî áåëêîâ òåïëîâîãî øîêà (HSP) ïðèíàäëåæèò ê ãðóïïå óáèêâèòàðíûõ ïåïòèäîâ, î÷åíü ÷óòêî ðåàãèðóþùèõ íà ôèçèîëîãè÷åñêèå è èììóíîëîãè÷åñêèå èçìåíåíèÿ â êëåòêå, ñïåöèàëèñòû ïîïûòàëèñü èñïîëüçîâàòü ýòè áåëêè äëÿ ïðîòèâîîïóõîëåâîé èììóíîòåðàïèè è èììóíîïðîôèëàêòèêè ìåòàñòàçîâ [67, 68]. Àâòîðû ïîëàãàþò, ÷òî ïîäîáíûé ïîäõîä, îñíîâàííûé íà ïîëó÷åíèè HSP èç îïóõîëåâûõ êëåòîê, â áóäóùåì ìîæåò îêàçàòüñÿ ïåðñïåêòèâíûì, óäîáíûì è íàäåæíûì ìåòîäîì ïîëó÷åíèÿ ïðîòèâîîïóõîëåâûõ âàêöèí.  ñâÿçè ñ òåì ÷òî ìíîãèå îïóõîëè ñèíòåçèðóþò â çíà÷èòåëüíîì êîëè÷åñòâå ïîëèñàõàðèäû, èìåþùèå ñèëüíûå àíòèãåííûå ñâîéñòâà, áûëà ïðåäïðèíÿòà ïîïûòêà [69] ïîëó÷èòü ñèíòåòè÷åñêèé ìîíîñàõàðèä â âèäå àëëèëãëèêîçèäà, àíàëîãè÷íîãî ïðèðîäíîìó àíòèãåíó Le, êîòîðûé çàòåì áûë êîíúþãèðîâàí ñ KLH.  äðóãîé ñåðèè èññëåäîâàíèé ïåíòàñàõàðèä áûë êîíúþãèðîâàí ñ ñûâîðîòî÷íûì àëüáóìèíîì. Ïîëó÷åííûé èììóíîãåí â ñî÷åòàíèè ñ àäúþâàíòîì QS-21 îêàçàëñÿ àêòèâíûì, ñóäÿ ïî ïðîäóêöèè çíà÷èòåëüíîãî êîëè÷åñòâà lgG è lgM, ÷òî äåëàåò åãî ïåðñïåêòèâíûì äëÿ èñïîëüçîâàíèÿ ïðè ðàçðàáîòêå ìåòîäîâ ïðîòèâîîïóõîëåâîé èììóíîòåðàïèè. Íà îñíîâå àíòèãåíîâ 17-1à, 791Ò gp72 è ÑÅÀ, ñïåöèôè÷åñêèõ äëÿ ðàêà êèøå÷íèêà, ðàçðàáîòàíû âàêöèíû, èíäóöèðóþùèå äîñòàòî÷íî ñèëüíûé èììóííûé îòâåò [68]. Ïðè ñðàâíåíèè ýôôåêòèâíîñòè ðàçëè÷íûõ âàêöèí, ñîçäàííûõ íà îñíîâå ÀÈÀÒ, ÄÍÊ-ôðàãìåíòîâ, ìóöèíîâ è áåëêîâ òåïëîâîãî øîêà, íå óñòàíîâëåíî çàìåòíûõ ïðåèìóùåñòâ êàêîé-ëèáî èç íèõ. Ýòî óêàçûâàåò íà òî, ÷òî íå òîëüêî ñàìà âàêöèíà, íî è äðóãèå ôàêòîðû, â ÷àñòíîñòè ðåæèì èììóíèçàöèè è ñâîéñòâà àäúþâàíòîâ, ìîãóò èìåòü ñóùåñòâåííîå çíà÷åíèå äëÿ ïîëó÷åíèÿ ìàêñèìàëüíîãî ïðîòèâîîïóõîëåâîãî ýôôåêòà. ÈÑÏÎËÜÇÎÂÀÍÈÅ ÊÎÌÏÎÍÅÍÒΠÃËÀÂÍÎÃÎ ÊÎÌÏËÅÊÑÀ ÃÈÑÒÎÑÎÂÌÅÑÒÈÌÎÑÒÈ (ÌÍÑ) Ðÿä èññëåäîâàíèé ïîñâÿùåí èñïîëüçîâàíèþ áåëêîâ ÌÍÑ ² èëè ²² êëàññà äëÿ ñîçäàíèÿ ïðîòèâîîïóõîëåâûõ âàêöèí [51, 7073]. ÌÍÑ ïðåä- 103 ñòàâëÿåò ñîáîé ñèñòåìó ãåíîâ, êîíòðîëèðóþùèõ ñèíòåç àíòèãåíîâ, îïðåäåëÿþùèõ íåñîâìåñòèìîñòü òêàíåé ïðè ïåðåñàäêå è èíäóöèðóþùèõ ðåàêöèè, âûçûâàþùèå îòòîðæåíèå òðàíñïëàíòàòîâ. Òàêàÿ îñîáåííîñòü ïðîäóêòîâ ýòèõ ãåíîâ ëåãëà â îñíîâó íîâûõ ïîäõîäîâ â ñîçäàíèè ïðîòèâîîïóõîëåâûõ âàêöèí. Ñ÷èòàåòñÿ, ÷òî ïðèñóòñòâèå áåëêîâ ÌÍÑ I êëàññà íà êëåòî÷íîé ïîâåðõíîñòè âåäåò ê óñêîëüçàíèþ êëåòîê îò ðàñïîçíàâàíèÿ èõ ýëåìåíòàìè èììóííîé ñèñòåìû. Ïðåäïîëàãàåòñÿ, ÷òî ïðèðîäíûå êèëëåðíûå êëåòêè ïîñòîÿííî ïðîâåðÿþò ñâîå ìèêðîîêðóæåíèå íà íàëè÷èå áåëêîâ ÌÍÑ ² êëàññà. Êëåòêè, óòðàòèâøèå òàêèå áåëêè, áóäóò ðàñïîçíàíû êàê ìèøåíè è óíè÷òîæåíû. B.A. Pulaski et al. [51] â ýêñïåðèìåíòàõ íà ìûøàõ ëèíèè BALB/c ñî ñïîíòàííûìè îïóõîëÿìè ìîëî÷íîé æåëåçû, ÿâëÿþùèìèñÿ áëèçêèì àíàëîãîì ðàêà ìîëî÷íîé æåëåçû ó ÷åëîâåêà ïî òàêèì ïîêàçàòåëÿì, êàê èììóíîãåííîñòü, ìåòàñòàòè÷åñêèå ñâîéñòâà è ðîñòîâûå õàðàêòåðèñòèêè, èììóíèçèðîâàëè æèâîòíûõ îïóõîëåâûìè êëåòêàìè, â êîòîðûå òðàíñôåöèðîâàëè ãåíû êëàññà ÌÍÑ ²² è îäíîâðåìåííî àíòèãåíû Â7-1.  ýòèõ îïûòàõ íàáëþäàëè ïîäàâëåíèå ðîñòà èëè ðàññàñûâàíèå ìåòàñòàçîâ, íî íå áûëî óñòàíîâëåíî âëèÿíèÿ èììóíèçàöèè íà ðîñò ïåðâè÷íîé îïóõîëè. Ýòè äàííûå ñâèäåòåëüñòâóþò, ÷òî òàêèå âàêöèíû, íàöåëåííûå íà àêòèâàöèþ CD4+- è CD8+-Ò-ëèìôîöèòîâ, ìîãóò áûòü ïðèìåíåíû â òåõ ñëó÷àÿõ, êîãäà ïåðâè÷íàÿ îïóõîëü õîðîøî ïîääàåòñÿ ëå÷åíèþ îáùåïðèíÿòûìè ìåòîäàìè, à åå ìåòàñòàçû óñòîé÷èâû ê òåðàïèè. Ýôôåêòèâíîñòü ïîëèâàëåíòíîé ïðîòèâîìåëàíîìíîé âàêöèíû, ïðèãîòîâëåííîé èç öåëüíûõ êëåòîê îïóõîëè, îêàçàëàñü îáóñëîâëåííîé ôåíîòèïè÷åñêèìè îñîáåííîñòÿìè ëåéêîöèòàðíîãî àíòèãåíà HLA I áîëüíîãî [70]. Ýòîò ôàêò ìîæåò óêàçûâàòü íà òî, ÷òî äëÿ âàêöèíîòåðàïèè ñëåäóåò îòáèðàòü áîëüíûõ ñ ñîîòâåòñòâóþùèì ôåíîòèïîì, à ó îñòàëüíûõ ïðèìåíÿòü äðóãèå âèäû ëå÷åíèÿ. CTL ðàñïîçíàþò ïåïòèäû íà ïîâåðõíîñòè îïóõîëåâûõ êëåòîê, ñâÿçàííûå ñ áåëêàìè ÌÍÑ ² êëàññà [74]. Ê ñîæàëåíèþ, îïóõîëåâûå àíòèãåíû, ïðåäñòàâëåííûå ëèáî êîäèðóåìûìè âèðóñíûì ãåíîìîì áåëêàìè, ëèáî áåëêàìè, âîçíèêàþùèìè â ïðîöåññå äèôôåðåíöèðîâêè, ëèáî, íàêîíåö, ìóòàíòíûìè áåëêàìè, çà÷àñòóþ ÿâëÿþòñÿ ñëàáûìè èììóíîãåíàìè. Ïîýòîìó ñêîíñòðóèðîâàíû ãåòåðîêëèòè÷åñêèå âàêöèíû, â êîòîðûõ ïåïòèäû ÌÍÑ ² êëàññà áûëè ñîåäèíåíû ñ àíòèãåíîì, à ÷åðåç ïåïòèäíûé ëèíêåð ñ ðåöåïòîðîì CTL [72]. Ðåçóëüòàòû èññëåäîâàíèé, âûïîëíåííûõ íà æèâîòíûõ ñ ìåëàíîìîé, ïîêàçàëè, ÷òî òàêîé ïîäõîä ìîæåò îêàçàòüñÿ ïåðñïåêòèâíûì â ñëó÷àå, êîãäà îïóõîëè ýêñïðåññèðóþò ñëàáîèììóíîãåííûå àíòèãåíû.  ïîñëåäíèå ãîäû ïîÿâëÿåòñÿ âñå áîëüøå ñâåäåíèé î òîì, ÷òî ìîëåêóëû HLA èãðàþò îãðîìíóþ ðîëü â ðàçâèòèè ïðîòèâîîïóõîëåâîãî èììóíèòåòà è ñïîñîáíîñòü îïóõîëåâûõ êëåòîê ïîäàâëÿòü ýêñïðåññèþ íåêîòîðûõ ìîëåêóë HLA èëè íàðóøàòü ñïåöèôèêó èõ âçàèìîäåéñòâèÿ ìîæåò áûòü îñíîâíûì ôàêòîðîì, ñíèæàþùèì ýôôåêòèâíîñòü èììóíîòåðàïèè. 104 Ýêñïåðèìåíòàëüíàÿ îíêîëîãèÿ 22, 99-109, 2000 (ñåíòÿáðü) ÈÑÏÎËÜÇÎÂÀÍÈÅ ÀÍÒÈÃÅÍÏÐÅÇÅÍÒÈÐÓÞÙÈÕ ÊËÅÒÎÊ (ÀÐÑ) Ïîñëåäíèå äåñÿòèëåòèÿ îçíàìåíîâàëèñü ðàñêðûòèåì ðÿäà ìîëåêóëÿðíûõ ìåõàíèçìîâ, îïðåäåëÿþùèõ ôîðìèðîâàíèå, ðàñïîçíàâàíèå, ïðîöåññèíã è àíòèãåííóþ ïðåçåíòàöèþ â èììóííîì îòâåòå. Óñòàíîâëåíèå ôàêòà, ÷òî ÀÐÑ èãðàþò âàæíåéøóþ ðîëü íå òîëüêî â ïðåçåíòàöèè, íî è â ïðåâðàùåíèè êîíôîðìàöèîííîãî ýïèòîïà àíòèãåíà â ëèíåéíûé, ñòèìóëèðîâàëî ïîÿâëåíèå ðàáîò, â êîòîðûõ ÀÐÑ èñïîëüçóþò â êà÷åñòâå ïðîòèâîîïóõîëåâûõ âàêöèí [2, 7579]. ÀÐÑ, íàãðóæåííûå íåôðàêöèîíèðîâàííûìè ïåïòèäàìè èç ãèäðîëèçàòà îïóõîëåâûõ êëåòîê, èíäóöèðîâàëè îòâåò CTL, óñòîé÷èâîñòü ê ïðèâèâêå îïóõîëåâûõ êëåòîê èëè ïîäàâëåíèå ìåòàñòàçèðîâàíèÿ [80]. Ð. De Matos et al. [75] âûäåëèëè äåíäðèòíûå êëåòêè èç êîñòíîãî ìîçãà ìûøåé C57Bl/6 ïóòåì êóëüòèâèðîâàíèÿ êëåòîê-ïðåäøåñòâåííèêîâ â ïðèñóòñòâèè ìûøèíîãî ãðàíóëîöèòàðíî-ìàêðîôàãàëüíîãî êîëîíèåñòèìóëèðóþùåãî ôàêòîðà (G-CSF). Èììóíèçàöèþ æèâîòíûõ îñóùåñòâëÿëè äåíäðèòíûìè êëåòêàìè, îáðàáîòàííûìè ëèçàòîì êëåòîê ìåëàíîìû  16, ïóòåì âíóòðèâåííûõ åæåíåäåëüíûõ èíúåêöèé. Òàêàÿ èììóíèçàöèÿ îáåñïå÷èâàëà çàäåðæêó îïóõîëåâîãî ðîñòà è óâåëè÷åíèå ñðåäíåé ïðîäîëæèòåëüíîñòè æèçíè ìûøåé. Ïðåäñòàâëÿë èíòåðåñ òàêæå òîò ôàêò, ÷òî èíäóêöèÿ ïðîòèâîîïóõîëåâîãî èììóíèòåòà áûëà ýôôåêòèâíà è ó æèâîòíûõ, êîòîðûì áûëà ïðèâèòà äðóãàÿ ãèñòîãåíåòè÷åñêè îòëè÷àþùàÿñÿ îïóõîëü ôèáðîñàðêîìà ÌÑÀ-106. Ýòè äàííûå ñâèäåòåëüñòâóþò î íàëè÷èè îáùèõ àíòèãåíîâ â êëåòêàõ ðàçëè÷íûõ ïî ãèñòîãåíåçó îïóõîëåé. Èñïîëüçîâàíèå äëÿ èììóíèçàöèè äåíäðèòíûõ êëåòîê, ïðîèñõîäÿùèõ èç êîñòíîãî ìîçãà, îáðàáîòàííûõ ëèçàòîì îïóõîëåâûõ êëåòîê, ïðèâîäèëî ê âîçíèêíîâåíèþ óñòîé÷èâîñòè ìûøåé ê ïîñëåäóþùèì ïðèâèâêàì ñìåðòåëüíîé äîçû îïóõîëåâûõ êëåòîê, è, ÷òî îñîáåííî âàæíî, ê âûðàæåííîìó ïîäàâëåíèþ ìåòàñòàçîâ â ëåãêèõ [77]. Ãèáåëü îïóõîëåâûõ êëåòîê çàâèñåëà îò êîëè÷åñòâà ÑD8 +- è â ìåíüøåé ñòåïåíè CD4+-Ò-ëèìôîöèòîâ. Âàêöèíàöèÿ äåíäðèòíûìè êëåòêàìè, íàãðóæåííûìè àíòèãåíàìè èç ëèçàòîâ êëåòîê ôèáðîñàðêîìû ÌÑÀ-106, ñïîñîáñòâîâàëà ïîäàâëåíèþ ìåòàñòàçèðîâàíèÿ îïóõîëåâûõ êëåòîê â ïå÷åíü [2]. Óäàëåíèå CD4+- è CD8+-Ò-ëèìôîöèòîâ ïîëíîñòüþ íèâåëèðîâàëî çàùèòíûé ýôôåêò âàêöèíû. Ïîêàçàíî, ÷òî äåíäðèòíûå êëåòêè ñåêðåòèðóþò àíòèãåíïðåçåíòèðóþùèå âåçèêóëû, íàçâàííûå ýêçîñîìàìè, êîòîðûå ñîäåðæàò áåëêè ÌÍÑ ² è ²² êëàññîâ, à òàêæå Ò-êëåòî÷íûå êî-ñòèìóëèðóþùèå ìîëåêóëû. Ýêçîñîìû, ïîëó÷åííûå ñ ïîâåðõíîñòè äåíäðèòíûõ êëåòîê, îêàçàëèñü ñïîñîáíûìè àêòèâèðîâàòü CTL, ÷òî ñîïðîâîæäàåòñÿ ïîëíîé ëèêâèäàöèåé èëè ïîäàâëåíèåì ðîñòà îïóõîëåé [1]. Àâòîðû ñ÷èòàþò, ÷òî âàêöèíû íà îñíîâå ýêçîñîì äåíäðèòíûõ êëåòîê ìîãóò ïðåäñòàâëÿòü õîðîøóþ àëüòåðíàòèâó àäîïòèâíûì âàêöèíàì íà îñíîâå öåëüíûõ äåíäðèòíûõ êëåòîê. Ìåòîäû ìîëåêóëÿðíîé áèîëîãèè è êëåòî÷íîé èíæåíåðèè ïîçâîëÿþò ñåãîäíÿ ìîäèôèöèðîâàòü îïóõîëåâûå êëåòêè òàê, ÷òî ïîâûøàåòñÿ èõ èììóíîãåííîñòü, àêòèâèðóþòñÿ ôóíêöèè CTL è äåíäðèòíûõ êëåòîê. Îñóùåñòâëåíû ïîïûòêè òðàíñäóêöèè ãåíîâ öèòîêèíîâ, êîñòèìóëèðóþùèõ ìîëåêóë èëè àëëîãåííûõ ìîëåêóë ÌÍÑ [81]. Ïðè ýòîì âîçíèêàåò óñòîé÷èâûé èììóíèòåò ê ïîñëåäóþùèì ïðèâèâêàì îïóõîëåâûõ êëåòîê.  1998 ã. áûëî îñóùåñòâëåíî ñâûøå 50 êëèíè÷åñêèõ èñïûòàíèé òàêîãî ìåòîäà èììóíîòåðàïèè [82]. Ïîëó÷åíèå äåíäðèòíûõ êëåòîê â çíà÷èòåëüíûõ êîëè÷åñòâàõ äëÿ ïðîòèâîðàêîâîé òåðàïèè â êëèíèêå ñòàëî ðåàëüíûì ïîñëå îïóáëèêîâàíèÿ ðàáîò [83, 84], â êîòîðûõ áûëî ïîêàçàíî, ÷òî èç ïðåäøåñòâåííèêîâ CD34+-êëåòîê ïåðèôåðè÷åñêîé êðîâè, ïîëó÷åííûõ ìåòîäîì ëåéêàôåðåçà, ïóòåì êóëüòèâèðîâàíèÿ â ïðèñóòñòâèè G-CSF/filgastrim â ñî÷åòàíèè ñ ôàêòîðîì íåêðîçà îïóõîëåé ìîãóò áûòü ïîëó÷åíû ïîëíîöåííûå äåíäðèòíûå êëåòêè, ìîðôîëîãè÷åñêè è èììóíîôåíîòèïè÷åñêè íå îòëè÷èìûå îò êëåòîê Ëàíãåðãàíñà êîæè, äåíäðèòíûõ êëåòîê êîñòíîãî ìîçãà è CD34+-êëåòîê èç ïóïî÷íîãî êàíàòèêà. Àêòèâèðîâàòü äåíäðèòíûå êëåòêè ìîæíî ïðè ïîìîùè äåçîêñèîëèãîíóêëåîòèäîâ, ñîäåðæàùèõ ôðàãìåíò ÑðG [85]. Âàæíî òàêæå, ÷òî ñïîñîáíîñòü äåíäðèòíûõ êëåòîê çàõâàòûâàòü, ïðîöåññèðîâàòü è ïðåçåíòèðîâàòü àíòèãåí çàâèñèò îò èñòî÷íèêà ïîëó÷åíèÿ äåíäðèòíûõ êëåòîê, à òàêæå îò ñòàäèè èõ ñîçðåâàíèÿ è äèôôåðåíöèðîâêè [84]. Äåíäðèòíûå êëåòêè ìîãóò áûòü íàãðóæåíû íå òîëüêî áåëêîâûì èëè ïîëèïåïòèäíûì àíòèãåíîì, íî è ñóììàðíîé êëåòî÷íîé ÐÍÊ, êîòîðàÿ ñ ïîìîùüþ ìåòîäîâ ãåííîé èíæåíåðèè ìîæåò áûòü íàðàáîòàíà â çíà÷èòåëüíîì êîëè÷åñòâå [86, 87]. Âàêöèíû, ïðèãîòîâëåííûå íà îñíîâå äåíäðèòíûõ êëåòîê, ïîëó÷åííûõ èç ëèìôîèäíûõ êëåòîê ïåðèôåðè÷åñêîé êðîâè, óæå ïðîøëè ïåðâóþ ïðîâåðêó â êëèíèêå ñ ïîëîæèòåëüíûì ýôôåêòîì [88]. Èíòåðëåéêèíû ÈË-7 è ÈË-10 çíà÷èòåëüíî óñèëèâàþò ñïîñîáíîñòü äåíäðèòíûõ êëåòîê çàïóñêàòü àíòèãåíñïåöèôè÷åñêèé îòâåò CTL. Ïðè ýòîì íàãðóæåííûå àíòèãåíîì äåíäðèòíûå êëåòêè ìîãóò áûòü ðàçìíîæåíû â êóëüòóðå â 100 èëè 1000 ðàç çà 1015 ñóò áåç ïîòåðè àêòèâíîñòè è ñïåöèôè÷íîñòè [89]. J. Cong et al. [90] ïîëó÷èëè ãèáðèäû äåíäðèòíûõ êëåòîê ñ êëåòêàìè êàðöèíîìû ÌÑ-38. Ââåäåíèå òàêèõ ãèáðèäíûõ êëåòîê æèâîòíûì âûçûâàëî àêòèâàöèþ CD4+- è CD8+-CTL, ÷òî çàùèùàëî èõ îò ïîñëåäóþùåé ïðèâèâêè îïóõîëåâûõ êëåòîê. Ââåäåíèå òàêèõ ãèáðèäíûõ êëåòîê æèâîòíûì ñ îïóõîëÿìè ïðèâîäèëî ê îòòîðæåíèþ óæå ðàçâèâøèõñÿ ìåòàñòàçîâ.  òî æå âðåìÿ îêàçàëîñü, ÷òî èììóíèçàöèÿ íåïîñðåäñòâåííî âûäåëåííûì îïóõîëåñïåöèôè÷åñêèì ïåïòèäîì â íåêîòîðûõ ñëó÷àÿõ ìîæåò âûçâàòü òîëåðàíòíîñòü CTL è äàæå ñòèìóëèðîâàòü îïóõîëåâûé ðîñò, ÷åãî íå íàáëþäàåòñÿ ïðè àäîïòèâíîé èììóíèçàöèè äåíäðèòíûìè êëåòêàìè [89, 91]. ÈÑÏÎËÜÇÎÂÀÍÈÅ ÍÓÊËÅÈÍÎÂÛÕ ÊÈÑËÎÒ Èñïîëüçîâàâøèåñÿ ðàíåå ñïîñîáû èììóíèçàöèè îñíîâàíû íà ââåäåíèè â îðãàíèçì æèâîòíîãî Experimental Oncology 22, 99-109, 2000 (September) èëè ÷åëîâåêà ñîîòâåòñòâóþùèõ àíòèãåíîâ. Îäíàêî â ïîñëåäíèå ãîäû âîçíèêëî íîâîå íàïðàâëåíèå, ñâÿçàííîå ñ ââåäåíèåì â îðãàíèçì íóêëåèíîâûõ êèñëîò (ÄÍÊ èëè ÐÍÊ), êîäèðóþùèõ ýòè áåëêè [68, 71, 84, 92, 93]. Áûëî ïîêàçàíî, ÷òî ââåäåíèå ïëàçìèäíîé ÄÍÊ â ìûøöó ìûøè ïðèâîäèò ê ïîãëîùåíèþ ýòîé ÄÍÊ êëåòêàìè è äëèòåëüíîé ýêñïðåññèè êîäèðóåìîãî ïëàçìèäîé âíåñåííîãî ãåíà [94]. Ïðîäîëæèòåëüíîñòü è èíòåíñèâíîñòü ñèíòåçà àíòèãåíîâ çàâèñèò îò ìåñòà ââåäåíèÿ ãåíåòè÷åñêîãî ìàòåðèàëà: ïðè ââåäåíèè â ñêåëåòíûå ìûøöû ñèíòåç ïðîäîëæàåòñÿ 36 ìåñ, â ñåðäå÷íóþ ìûøöó 14 60 ñóò [9598]. Ââåäåííàÿ ÄÍÊ íå âñòðàèâàåòñÿ â ãåíîì, à ñóùåñòâóåò â âèäå ýïèñîìû ïðîäîëæèòåëüíîå âðåìÿ. Ðàçðàáîòàí ðÿä ñïîñîáîâ ââåäåíèÿ ÄÍÊ â êëåòêè. Ïðåäëîæåí áàëëèñòè÷åñêèé ìåòîä, ïðè êîòîðîì ìèêðîøàðèêàìè çîëîòà, ïîêðûòûìè ÄÍÊ, áîìáàðäèðóþò ïîâåðõíîñòü êëåòîê ïðè ïîìîùè ãåííîé ïóøêè [99, 100]. Ýòîò ìåòîä óñïåøíî ïðèìåíåí ïðè ëå÷åíèè áîëüíûõ c ìåëàíîìîé è äðóãèìè îïóõîëÿìè êîæè [101]. Ïî-âèäèìîìó, äëÿ âñòðàèâàíèÿ ÄÍÊ â êëåòêè-ìèøåíè ìîæåò áûòü èñïîëüçîâàí è ìåòîä ýëåêòðîïîðàöèè [102].  ïðîñòåéøèõ ñëó÷àÿõ ÄÍÊ ìîæåò áûòü ââåäåíà òîíêîé èãëîé âíóòðèêîæíî, ëèáî ìåòîäîì ñêàðèôèêàöèè [71]. Óñèëåíèå ñèíòåçà áåëêîâ Â7 [103], áåëêîâ ÌÍÑ ² êëàññà [104], öèòîêèíîâ [105, 106] ïîñëå ââåäåíèÿ ñîîòâåòñòâóþùèõ ãåíîâ ñïîñîáñòâóåò èíäóêöèè êëåòî÷íîãî ïðîòèâîîïóõîëåâîãî èììóíèòåòà. ÄÍÊ ìîæåò áûòü ââåäåíà â ñîñòàâå êàòèîííûõ èëè ðÍ-çàâèñèìûõ ëèïîñîì, à òàêæå â ñîñòàâå ðåêîìáèíàíòíûõ âèðóñîâ, äëÿ ÷åãî îáû÷íî èñïîëüçóþò ðåòðî- [107] è àäåíîâèðóñû [108]. Ìèøåíÿìè äëÿ ïðîòèâîîïóõîëåâîãî èììóííîãî îòâåòà ìîãóò ñëóæèòü ãëèêîïåïòèäû, ãëèêîëèïèäû, ãëèêîïðîòåèäû, àíòèãåíû äèôôåðåíöèðîâêè (MAGE, òèðîçèíàçà, ÌÅËÀÍÀ, gp-75), ïðîäóêòû îíêîãåíîâ (ð53, HER2/neu) è äðóãèå, ñèíòåç êîòîðûõ ìîæåò áûòü çàïóùåí ââåäåíèåì ñîîòâåòñòâóþùèõ ïîñëåäîâàòåëüíîñòåé ÄÍÊ. Âàêöèíàöèÿ ìûøåé ðåêîìáèíàíòíûì âèðóñîì, ýêñïðåññèðóþùèì íîðìàëüíûé áåëîê ð53, çàùèùàåò æèâîòíûõ îò ðàçâèòèÿ îïóõîëåé èç òðàíñôîðìèðîâàííûõ ôèáðîáëàñòîâ [109]. Ïåðñïåêòèâíûì ìîæåò îêàçàòüñÿ îäíîâðåìåííîå ââåäåíèå íåñêîëüêèõ âåêòîðîâ, êîäèðóþùèõ îïóõîëåâûå àíòèãåíû, ãåíû àïîïòîçà è öèòîêèíîâ [71]. Ñîâìåñòíîå ââåäåíèå â îïóõîëåâûå êëåòêè ãåíîâ òèìèäèíêèíàçû è ÈË-2 ïðèâîäèò ê ðåãðåññèè íîâîîáðàçîâàíèÿ è ôîðìèðîâàíèþ ïðîòèâîîïóõîëåâîãî èììóíèòåòà [110]. Äëÿ ñîçäàíèÿ ïðîòèâîîïóõîëåâîãî èììóíèòåòà èñïîëüçóþò òàêæå ìåõàíèçìû, çàïóñêàþùèå èëè áëîêèðóþùèå àïîïòîç. Íàïðèìåð, ââåäåíèå â îïóõîëü MCF-7 âåêòîðà íà îñíîâå àäåíîâèðóñà ñ ó÷àñòêîì ÄÍÊ, êîäèðóþùèì ðåïðåññîð áåëêîâ Bcl-2 è Bcl-x(L), ïðèâîäèò ê ðåãðåññèè 50% îïóõîëåé ó ìûøåé [111]. Íåêîòîðûå àâòîðû, îäíàêî, ïðåäóïðåæäàþò î âîçìîæíûõ íåæåëàòåëüíûõ ïîñëåäñòâèÿõ, ñâÿçàííûõ ñ ââåäåíèåì â îðãàíèçì ÷óæåðîäíûõ íóêëåèíîâûõ êèñëîò [112]. 105 ÄÐÓÃÈÅ ÏÎÄÕÎÄÛ Äëÿ ïîâûøåíèÿ èììóííîãåííîñòè àíòèãåíîâ ïðåäëîæåíî ïðè èçãîòîâëåíèè âàêöèí èñïîëüçîâàòü èçîëèðîâàííûå ýíäîñîìû è ýêçîñîìû ÀÐÑ, ïîñêîëüêó â ýòèõ ñòðóêòóðàõ îñóùåñòâëÿåòñÿ àññîöèàöèÿ àíòèãåíà ñ áåëêàìè ÌÍÑ ²² êëàññà [113]. Ïîêàçàíî, ÷òî òàêèå âàêöèíû ñïîñîáíû ñòèìóëèðîâàòü Ò-êëåòêè in vitro. Êðîìå òîãî, ýêçîñîìû áîãàòû êî-ñòèìóëèðóþùèìè ìîëåêóëàìè CD86, CD37, CD53, CD63, CD81, CD82. Ðàçðàáîòàí ìåòîä äîëãîñðî÷íîãî êóëüòèâèðîâàíèÿ îïóõîëåâûõ êëåòîê ÷åëîâåêà, à òàêæå èõ ñìåñè ñ ëèìôîöèòàìè äëÿ àäîïòèâíîé èììóíîòåðàïèè [114]. L. Kwak et al. [115] ïðåäëîæèëè èñïîëüçîâàòü èììóíîãëîáóëèí Ì èç êëåòîê ëèìôîìû ìûøåé 38Ñ13 äëÿ èììóíèçàöèè â ôîðìå ëèïîñîì.  ýòèõ îïûòàõ îäíîêðàòíàÿ èììóíèçàöèÿ îáåñïå÷èâàëà çàùèòó æèâîòíûõ ïðè ââåäåíèè ñìåðòåëüíîé äîçû îïóõîëåâûõ êëåòîê. Ýôôåêòèâíîñòü âàêöèíû âîçðàñòàëà ïðè äîïîëíèòåëüíîì ââåäåíèè ÈË-2. Ïîêàçàíî, ÷òî ââåäåíèå öèêëîôîñôàìèäà ïîñëå àêòèâíîé ñïåöèôè÷åñêîé èììóíèçàöèè ïîâûøàåò ïðîòèâîîïóõîëåâûé èììóíèòåò, ÷òî ñâÿçûâàåòñÿ ñ ïîäàâëåíèåì êëåòîê-ñóïðåññîðîâ â ñîñòàâå CD4+T-ëèìôîöèòîâ [116]. Ïðåäëîæåíà âàêöèíà, ïðè èììóíèçàöèè êîòîðîé ïîìèìî àêòèâàöèè ëèìôîèäíûõ êëåòîê èíäóöèðóåòñÿ ñèíòåç êîëëàãåíîâûõ âîëîêîí [117]. Ïðè ýòîì êîëëàãåí IV òèïà îáðàçóåòñÿ âíóòðè îïóõîëè, à êîëëàãåíû ² è ²² òèïà, à òàêæå ôèáðîíåêòèí âîêðóã îïóõîëåâîãî óçëà. Ïîñëå èììóíèçàöèè íàáëþäàëè èíêàïñóëèðîâàíèå îïóõîëè, ÷òî ïðèâîäèëî ê ïîäàâëåíèþ åå ðîñòà è ìåòàñòàçèðîâàíèÿ. Îïèñàí ïîëîæèòåëüíûé ýôôåêò ïðè èììóíèçàöèè êóð ðåêîìáèíàíòíûì âèðóñîì êóðèíîé îñïû (rFPV-gp), êîäèðóþùèì ñèíòåç â êëåòêàõ ñåëåçåíêè ãëèêîïåïòèäà, õàðàêòåðíîãî äëÿ âèðóñà áîëåçíè Ìàðåêà. Ïîñëå âàêöèíàöèè ñíèæàëñÿ ïðîöåíò çàáîëåâøèõ ïòèö ïðè èíôèöèðîâàíèè îíêîãåííûì âèðóñîì [118].  ñâÿçè ñ òåì ÷òî ìåìáðàíû îïóõîëåâûõ êëåòîê îòëè÷àþòñÿ ïîíèæåííîé ìèêðîâÿçêîñòüþ, è ýòà èõ îñîáåííîñòü â çíà÷èòåëüíîé ìåðå îïðåäåëÿåòñÿ ñíèæåíèåì óðîâíÿ õîëåñòåðèíà, ïðåäïðèíÿòû ïîïûòêè èñïîëüçîâàòü äëÿ èììóíèçàöèè ìåìáðàíû îïóõîëåâûõ êëåòîê ñ èñêóññòâåííî ïîâûøåííûì ñîäåðæàíèåì õîëåñòåðèíà. Ââåäåíèå â ñòðóêòóðó ìåìáðàí õîëåñòåðèíà ãåìèñóêöèíàòà ñîïðîâîæäàëîñü ïîâûøåíèåì ìèêðîâÿçêîñòè ìåìáðàí è îäíîâðåìåííî ïîâûøåíèåì èõ èììóíîãåííîñòè [119121]. Ýôôåêòèâíîñòü òàêèõ âàêöèí ïîêàçàíà â îïûòàõ íà õîìÿêàõ ñ êàðöèíîìîé ïîäæåëóäî÷íîé æåëåçû [119] è ó ìûøåé ñ îïóõîëüþ ìîëî÷íîé æåëåçû è Ò- è Â-ëèìôîìàìè [121]. Çàòåì èììóíèçàöèè áûëè ïîäâåðãíóòû áîëüíûå ñ ñîëèäíûìè îïóõîëÿìè, êîòîðûì ââîäèëè âíóòðèìûøå÷íî ñ äâóõíåäåëüíûì èíòåðâàëîì îáëó÷åííûå è îáðàáîòàííûå õîëåñòåðèíîì àóòîëîãè÷íûå îïóõîëåâûå êëåòêè. Ó 7 èç 21 áîëüíîãî íàáëþäàëè ðåãðåññèþ îïóõîëåâûõ óçëîâ [120]. Ãðóïïà èññëåäîâàòåëåé èç Âåéöìàíîâñêîãî èíñòèòóòà (Èçðàèëü) [122, 123] îáíàðóæèëà, ÷òî ñêðûòûå îïóõîëåâûå àíòèãåíû è áåëêè ÌÍÑ íà ïîâåðõíîñòè îïóõîëåâûõ êëåòîê îáíàæàþòñÿ ïîä âëèÿíèåì âû- 106 Ýêñïåðèìåíòàëüíàÿ îíêîëîãèÿ 22, 99-109, 2000 (ñåíòÿáðü) ñîêîãî ãèäðîñòàòè÷åñêîãî äàâëåíèÿ. Íà êëåòêè ìåëàíîìû Â16 âîçäåéñòâîâàëè äàâëåíèåì 1200 àòì â òå÷åíèå 15 ìèí, à çàòåì ñøèâàþùèì àãåíòîì àäåíîçèí-2'-3'-äèàëüäåãèäîì. Èììóíîãåííîñòü ìîäèôèöèðîâàííûõ òàêèì îáðàçîì êëåòîê çíà÷èòåëüíî âîçðàñòàëà, ÷òî ñîïðîâîæäàëîñü ïîâûøåíèåì àêòèâíîñòè CTL è åñòåñòâåííûõ êèëëåðíûõ êëåòîê. Àâòîðû ñ÷èòàþò, ÷òî òàêèì ñïîñîáîì ìîãóò áûòü ïðèãîòîâëåíû ïðîòèâîîïóõîëåâûå âàêöèíû, ïðèãîäíûå äëÿ êëèíè÷åñêîãî ïðèìåíåíèÿ. Ïîâûøåíèå àêòèâíîñòè ïðîòèâîîïóõîëåâûõ âàêöèí ìîæåò áûòü äîñòèãíóòî èñïîëüçîâàíèåì íîâûõ, áîëåå ýôôåêòèâíûõ àäúþâàíòîâ, ÈË [124], ìèêðîýëåìåíòîâ [125, 126] è ïðåïàðàòîâ ïðèðîäíîãî ïðîèñõîæäåíèÿ. ÇÀÊËÞ×ÅÍÈÅ Ïðèâåäåííûå äàííûå ïîêàçûâàþò, ÷òî, íåñìîòðÿ íà äëèòåëüíóþ èñòîðèþ, ðàçðàáîòêà ïðîòèâîîïóõîëåâûõ âàêöèí íàõîäèòñÿ íà íà÷àëüíîì ýòàïå. Èññëåäîâàíèÿ â îáëàñòè ñîçäàíèÿ ïðîòèâîîïóõîëåâûõ âàêöèí âåäóòñÿ â ðàçëè÷íûõ íàïðàâëåíèÿõ è èõ êîëè÷åñòâî óâåëè÷èâàåòñÿ. Èìåþùèåñÿ ðåçóëüòàòû íå ïîçâîëÿþò ñäåëàòü îêîí÷àòåëüíûé âûâîä î ïðåèìóùåñòâå êàêîãî-ëèáî íàïðàâëåíèÿ. Äîñòèæåíèÿ ìîëåêóëÿðíîé áèîëîãèè è êëåòî÷íîé èíæåíåðèè îòêðûëè íîâûå ïîäõîäû ê ñîçäàíèþ ýôôåêòèâíûõ èììóíîãåíîâ è íåñïåöèôè÷åñêèõ ñòèìóëÿòîðîâ ïðîòèâîîïóõîëåâîãî èììóííîãî îòâåòà, à òàêæå âîçìîæíîñòè ìîíèòîðèíãà íàïðÿæåííîñòè îòäåëüíûõ çâåíüåâ èììóííûõ ðåàêöèé. 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