Isingeniso
I-CAR T cell therapy, uhlobo olusha lwe-immunotherapy, ibonise ukusebenza kahle okukhethekile ekwelapheni izimila ezithile ngokusebenzisa amandla amasosha omzimba esiguli. Muva nje, ososayensi baphenye umqondo wokuyisebenzisa ukwelapha izifo zamaseli B ahambisana ne-HIV, okuyinkinga eyinkimbinkimbi ehlanganisa kokubili umdlavuza kanye ne-HIV/AIDS.
Iziguli ezine-HIV zinengozi yomdlavuza ephakeme kakhulu kunesibalo sabantu esivamile. Abantu abane-HIV banamazinga omdlavuza angama-25-40%. Isifo esivame kakhulu ezigulini ezine-HIV/AIDS I-non-Hodgkin lymphoma (NHL). NHL is most common in PLWHA as Burkitt lymphoma, which has an 18-fold higher risk, and diffuse large B i-cell lymphoma (DLBCL), which accounts for 75% of cases.
I-Chimeric antigen receptor (CAR) T cell therapy, especially CD19-targeting CAR T cells, has revolutionized B cell NHL treatment. Due to safety concerns and the difficulties of generating CAR T cell products from HIV-positive donors, PLWHA has been excluded from all clinical studies for years. Recent studies investigated the use of HIV-positive i-lymphoma donors’ cells to create CD19-targeting CAR T cells. These studies have shown promise in addressing the safety and efficacy of Ukwelashwa kwe-CAR T cell kulesi sibalo sabantu, kodwa abaphathi HIV.
In the past, CD19-CAR and N6-CAR T cells were effective in both clinical and preclinical testing against lymphoma and HIVgp120. It was anticipated that a dual design targeting both antigens may target cancer and HIV-infected cells in one person. A bi-specific N6-huCD19 CAR T cell technology was developed that targets both antigens in one therapeutic product.
Ukuqonda inselele
Abantu abane-HIV/AIDS banethuba eliphezulu lokuthola izinhlobo ezithile zomdlavuza, ikakhulukazi ama-B cell malignancies okuhlanganisa i-lymphomas kanye ne-leukemia. Ukwelapha lezi zimila lapho kukhona i-HIV kungase kube inselele ngenxa yokuntenga kokuzivikela komzimba kanye nobunzima bokulawula ukucindezelwa kwamasosha omzimba.
Isebenza kanjani i-CAR T therapy?
I-CAR (chimeric antigen receptor) Ukwelashwa kwe-T cell ukuguqulwa kofuzo kwamaseli T esiguli ukuze aveze ama-receptor okwenziwa, abizwa ngokuthi ama-CAR, ebusweni bawo. Lezi zimoto zenzelwe ukuhlonza izinto ezithile ezitholakala kumaseli ayingozi. Phakathi kwezifo ezithinta amaseli e-B ahlobene ne-HIV, amaseli e-CAR T aklanyelwe ngokukhethekile ukubona ama-antigen afana ne-CD19, avame ukuba khona kuma-B cell.
Izinzuzo nezinselele Ezigulini ezine-HIV
Ukusebenzisa ukwelashwa kwe-CAR T cell kubantu abane-HIV abane-B cell malignancies kunikeza izinzuzo nezithiyo ezihlukene. Inzuzo yamaseli e-CAR T ukucaciswa kwawo okuphezulu ekuqondiseni amangqamuzana omdlavuza kuyilapho elondoloza amaseli anempilo, okungahle anciphise ingozi yobuthi. Noma kunjalo, ukutheleleka nge-HIV kubangela izinselele ekusebenzeni kwamasosha omzimba, okudinga ukuphathwa ngokucophelela kwakho kokubili igciwane lesandulela ngculaza kanye nesifo esibulalayo.
Ukuthuthukiswa kwe-CAR-T ye-HIV
ISIGABA 1. Ukumelwa okuhleliwe kwesakhiwo se-CAR esilwa ne-HIV. I-CAR yesizukulwane sokuqala siqukethe isizinda se-env esisodwa esilwa ne-HIV (i-CD4 noma i-svFc), isizinda sesizinda se-transmembrane, kanye nesizinda se-T cell receptor CD3-ζ. I-CAR yesizukulwane sesibili ihlanganisa isizinda esingeziwe sokubonisa i-costimulatory ekucushweni okuyisisekelo kwe-receptor yesizukulwane sokuqala. Amaseli esizukulwane sesithathu aqukethe isizinda esingaphezu kwesisodwa esikhuthazayo. Ama-CAR esizukulwane sesine abonakala ngokungezwa kwama-transgene ayisisekelo noma angaguquki njengama-cytokines noma ama-chemokines.
Kwakhiwe amadizayini ekhandidethi amabili e-tandem kanye neluphu eyodwa eqondene ne-CAR. Lezi zakhiwo zidalwe ngokuhlanganisa i-CD19 ye-CD6 eyodwa-chain fragment (scFv) kanye ne-N2 scFv ephuma ku-anti-HIV ebanzi evimbela i-HIV (bNAb) ibe umgogodla wokwakha we-CAR wesizukulwane sesibili.
The bi-specific tandem CAR constructs had N6 and huCD19 scFvs coupled with a G4S linker in huCD19-N6 or N6-huCD19 orientation. The loop-CAR was created by fusing huCD19(VL):N6(VH):VL:VH with a whitlow linker. All constructs had the CD4 transmembrane domain, a double-mutated IgG4 Fc spacer, 4-1BB co-stimulatory and CD3-zetta signaling domains, and EGFRt separated by a T2A ribosomal skip region. The 3 bi-specific constructs were assessed in healthy donor-derived T cells employing cytotoxicity co-culture assay against Raji (CD19+) or 8E5 (gp120+) target cells. N6-huCD19 CAR T cells were produced from HIV-positive patients and tested against isisu amaseli.
Imiphumela
Ukuhlola okusebenzayo kubonise ukuthi yomithathu imiklamo ye-CAR ene-bi-specific yayisebenza ngempumelelo ku-CD19 ne-HIVgp120. I-N6-huCD19 tandem CAR T cell yayisebenza kangcono ngokumelene nawo womabili ama-antigen. Ngokuphawulekayo, i-N6-huCD19 tandem CAR T amaseli angase aqonde ngokulandelana kwe-antigen eyodwa noma womabili.
I-N6-huCD19 tandem CAR T amaseli zakhiwe kubanikeli abane-HIV, okufakazela ukuthi le ndlela ingenzeka ku-PLWHA. Kuyathakazelisa ukuthi amaseli e-CAR T asuselwa kumnikeli we-HIV abulala amaseli e-tumor ayeveza ama-CD6 noma ama-antigen e-HIVgp19, okubonisa ukuthi angenza izinto ezimbili ezihlukene.
Ungahle uthande ukufunda: I-CAR T Cell therapy eShayina
ISIGABA 2. I-Anti-HIV CAR iguqule ama-T cell kanye nesu elithi “Kick and Kill” lokuqeda amaseli asanda kutheleleka nge-HIV. Isu “lokukhahlela” lisebenzisa ama-LRA ukuheha ukubhalwa kwe-HIV, ukuvezwa kwamaprotheni, nokukhiqizwa kwe-virion. I-CD4 (phezulu kwesokunxele, iwolintshi) noma i-antibody (phezulu kwesokudla, onsomi)-based CAR eyenziwe ngobunjiniyela iqondise indawo ebopha i-HIV ezinhlayiyeni zegciwane endaweni yeseli yereservoir cell evuselelwe kabusha (ngezansi). Ngemva kokubopha, i-T cell elungiswe yi-CAR izokhipha ama-granzyme nama-cytokines ukuze "abulale" amaseli ane-HIV.
Ukubhekana ne-Immunocompromise
An essential factor to address in Ukwelashwa kwamaseli e-CAR T for HIV-related malignancies is the equilibrium between cancer treatment and HIV control. Individuals with weakened immune systems may be more prone to infections and may also face potential problems from immunotherapy treatment. Hence, it is imperative to closely evaluate and implement tailored treatment options.
Amaseli E-CAR T Okulwa Nokutheleleka Nge-HIV
Ukusetshenziswa kwama-T cell okutholwa ukwelapha i-HIV kwakhuthazwa emashumini eminyaka edlule. Uhlolo lokuqala lwe-HIV CAR T cell therapy lwalubandakanya ukufakelwa kabusha kwama-T cell okutholwa nge-CAR fusion yesizinda se-CD4 extracellular (i-HIV receptor eyinhloko) kusizinda sokusayina se-CD3 (CD4) (Mitsuyasu et al., 2000; Walker et al., 2000; Deeks et al., 2002). I-CD4, ingxenye yemvelo yokuqaphela imvilophu ye-HIV, iqondise zonke izinhlobo ze-HIV, okuyenza ibe i-ligand esebenza kahle yokuklama i-CAR elwa ne-HIV. Ukuguqulwa kwezakhi zofuzo kunganciphisa ukubopha kwe-CD4 kanye nokufaneleka kwegciwane, yingakho, amaprotheni emvilophu e-CD4 amasayithi abophayo ayalondolozwa kakhulu (Wang et al., 2019).
I-CAR yesizukulwane sokuqala esekwe ku-CD4 yahlolwa ezigulini ezine-HIV ukuze isebenze kahle nokuphepha (Mitsuyasu et al., 2000; Walker et al., 2000; Deeks, 2002). Ukwelashwa akuzange kuvimbele ukuphindaphinda kwegciwane, kodwa amaseli ashintshiwe ahlala> iminyaka engu-10 futhi ayengenabo ubuthi obucacile (Mitsuyasu et al., 2000). Ukuntuleka kokulawulwa kwegciwane kungase kube nezimbangela eziningi: (1) Ama-CAR asekelwe ku-CD4 enza ama-T cell ashintshwe izakhi azwele ekuthelelekeni nge-HIV nokufa kwamangqamuzana, (2) isignali yokuvula isignali ye-costimulatory ingasebenzi kahle, (3) ukuphatha nokuphindaphinda kwamaseli e-T akwanele. impofu, futhi (4) ukukhuthazwa kwe-antigen yegciwane akukho. Kodwa-ke, ukwelashwa kwe-CD4ζ T cell kutholwe kuphephile futhi kugcinwe amazinga okufakwa ezinzile (Scholler et al., 2012).
Ungahle uthande ukufunda: Ukwelashwa kwe-CAR T Cell eNdiya
Intuthuko yakamuva ekwakhiweni kwe-CAR yokuthuthukisa ukusebenza kweseli ye-CAR T nokuphikelela ebubini kusheshise ukwelashwa kwamaseli e-CAR T. Izizukulwane ezine zama-CAR zikhona (Umfanekiso 1). Isizukulwane sokuqala sama-CAR sixhumanise i-antigen yangaphandle yokuqaphela i-moiety ku-endodomain ye-lymphocyte-stimulating intracellular, njengengxenye ye-TCR CD3ζ ye-signal-transducing component (Eshhar et al., 1993).
I-Apoptosis, elinganiselwe ekukhuleni kwe-vivo, kanye ne-cytotoxicity yayivamile kumaseli e-CAR T esizukulwane sokuqala (Heuser et al., 2003; Zhao et al., 2009). Ama-CAR esizukulwane sesibili adalwe ngokungeza izizinda ze-molecule ezibizayo njenge-CD28 noma i-4-1BB kumsila we-cytoplasmic wezakhiwo eziqukethe i-CD3ζ. I-In vitro, amaseli e-CAR T esizukulwane sesibili alwa ne-HIV anesizinda se-costimulatory 4-1BB acindezele ukuphindaphinda kwe-HIV ngokuphindwe ka-50 kangcono kunamaseli e-CAR T esizukulwane sokuqala (Leibman et al., 2017).
Ukuhlolwa kwezilwane kubonise ukuthi ama-CAR esizukulwane sesibili ande ekuphenduleni i-antigen, avikela amaseli e-CD4 + T ekuthelelekeni nge-HIV, futhi anciphisa ukuncipha kwe-CD4 kangcono kunama-CAR ngaphandle kwama-molecule e-costimulatory (Leibman et al., 2017). Ucwaningo oluqhathaniswayo lubonise ukuthi isizinda se-costimulatory se-4-1BB sinciphisa ukuphindaphinda kwegciwane ngemva kokwelashwa kwe-ART futhi sikhuthaza ukuphikelela kwe-T cell ku-vivo ngaphandle kwe-antigen kangcono kunesizinda se-CD28 (Zhang et al., 2007; Leibman et al., 2017). Ama-CAR esizukulwane sesithathu akhiqizwa ngokungeza ama-molecule amaningi e-costimulatory kuma-CAR esibili. Kumdlavuza, ama-CAR esizukulwane sesithathu athuthukisa ukusebenza kahle, ukwanda, ukusinda, nokufa kwe-tumor (Savoldo et al., 2011).
Liu et al. (I-2016) ithole ukuthi isizukulwane sesithathu se-anti-gp120 CAR enezizinda eziningi zokusayina ze-intracellular (CD3ζ-CD28-41BB) inempumelelo enkulu ekuqambeni amaseli e-Env-expressing in vitro kune-CD4ζ-CAR. Amaseli e-CAR T esizukulwane sesine, aziwa ngokuthi ama-T cell aqondiswe kabusha ekubulaweni kwe-cytokine-mediated universal (TRUCKs), aqukethe isignali yesithathu evuselelayo efihla noma ebopha ama-cytokines afana ne-IL-7, IL-12, IL-15, noma IL-18 ukuze ngcono ukunwetshwa kwamaseli e-CAR T nokuphikelela. Bakhona wafunda ku-oncology ukukhomba izimila eziqinile.
Isiphetho
Ukwelashwa kwe-CAR T cell kunamandla amakhulu njengendlela yokwelapha entsha ehlobene ne-HIV yamaseli e-B, okunikeza ithemba elisha kubantu ababhekene nalesi sifo esiyinkimbinkimbi. Naphezu kokuba khona kobunzima, ucwaningo oluqhubekayo kanye nemizamo yokwelashwa kuholela ekuthuthukisweni kokwelashwa okuthuthukisiwe nokulungiselelwe okunamandla okuguqula ukunakekelwa komdlavuza kulabo abaphila ne-HIV.
Miningi imizamo eyenziwayo yokwakha amasu amasha okuqeda i-HIV. Amasu asekelwe e-CAR angasiza ekuqedeni i-HIV ngokuthuthukisa impendulo yokuzivikela kumaselula elwa namagciwane. Inhlanganisela yamasu angase adingeke ukuze kusetshenziswe ngempumelelo ukwelashwa kwe-CAR okulwa ne-HIV ukuze kusungulwe ukubhekwa komzimba wesikhathi eside nokuqeda amaseli e-HIV avuselelwe. Kodwa-ke, eminye imibuzo isasele mayelana nokuthuthuka kwalo mkhakha:
(1) Ingabe amaseli omzimba ashintshwe i-CAR angakwazi ukubona inkulumo ye-antigen eyenziwe yasebenza kabusha ye-LRA?
(2) Ingabe ukwelashwa kwe-CAR kanye nama-LRA kungathinta amathumbu namathangi ezicubu zobuchopho?
(3) Ingabe i-CAR-modified immune cell infusions kanye/noma ukuvuselelwa kwe-LRA kuyadingeka?
Ukwelashwa kweseli ye-CAR T kwesizukulwane esilandelayo njengengxenye yohlelo “lokukhahlela nokubulala”, kuhlanganiswe nama-LRA noma ama-bNAbs, kungase kuqede indawo yokugcina i-HIV ngokuthuthukisa ukubhekwa kwamasosha omzimba nokugcina ukucindezelwa kwegciwane ngemva kokuphazanyiswa kwe-ART.
