Research ArticleExperimental Studies
Open Access

The Expression of LDL-R in CD8+ T Cells Serves as an Early Assessment Parameter for the Production of TCR-T Cells

YI WANG, YA-NAN HAO, MEI-YING SHEN, XIAO-JIAN HAN, YING-MING WANG, SI-YIN CHEN, JUN-FAN WANG, WANG WANG, TING-TING LI and AI-SHUN JIN
In Vivo November 2023, 37 (6) 2480-2489; DOI: https://doi.org/10.21873/invivo.13355

Abstract

Background/Aim: The quantity and the phenotypes of desired T cell receptor engineered T (TCR-T) cells in the final cell product determine their in vivo anti-tumor efficacy. Optimization of key steps in the TCR-T cell production process, such as T cell activation, has been shown to improve cell quality. Materials and Methods: Using a modified TCR (mTCR) derived from mice transducing PBMCs, we assessed the proportions of low-density lipoprotein receptor (LDL-R) and mTCR expressing cells under the various activation conditions of CD3/CD28-Dynabeads or OKT3 via flow cytometry. Results: We demonstrate that the proportion of T cells expressing LDL-R post activation is positively correlated with the percentage of mTCR+CD8+ T cells with their less differentiated subtypes in the final product. In addition, we show that shifting the CD3/CD28-Dynabeads activation duration from a typical 48 h to 24 h can significantly increase the production of the desired mTCR+CD8+ T cells. Importantly, the percentages of TCR-T cells with less-differentiated phenotypes, namely mTCR central memory T cells (TCM), were found to be preserved with markedly higher efficiency when T cell activation was optimized. Conclusion: Our findings suggest that the proportion of LDL-R+ T cells may serve as an early assessment parameter for evaluating TCR-T cell quality, possibly facilitating the functional and economical improvement of current adoptive therapy.

Key Words:

The adoptive transfer of TCR-T cells expressing tumor-specific antigens is a promising approach for cancer treatment (1-5). Within the multi-step process of TCR-T cell production, T cell activation is a critical event that has been implicated in determining the success rate of genetic modification and the quality of the desired proportion of T cell product (2, 3, 6). In specific, the susceptibility of T cells to lentiviral transduction is shown to be influenced by the stimulating approach, which has been a focused area of optimization study for the application of adoptive therapy (7-10).

Among the prevailing activation methods of human T cells, plate-coated anti-CD3 antibody (OKT3) and soluble anti-CD28 antibody are the first commercially available approaches, with affordability and practical simplicity (11-14). A later developed activation method using cell-sized Dynabeads coated with anti-CD3 and anti-CD28 antibodies (CD3/CD28-Dynabeads) possesses comparatively better efficiency (8, 15, 16). At present, typical T cell activation markers, including CD69 and CD25, were employed as indicators of T cell activation status (9, 17). However, very few parameters were applicable to effectively predict the proportion and the phenotypes of TCR-T cells in the final cell product of adoptive transfer. Therefore, identification of capable factors evaluating TCR-T cell quality early as during T cell activation would be fundamental for the strategical improvement of the production process.

LDL-R serves as a major entry receptor for the fusiogenic envelope G glycoprotein of the vesicular stomatitis virus, which is used to produce lentivirus or retrovirus vectors for gene transfer (18). The expression of LDL-R has been shown to be profoundly elevated upon T cell activation (19, 20). Augmented LDL-R on T cell surface facilitates the lentiviral recognition, which, in turn, determines the transduction efficiency of the desired TCR (21). Whether the ratio of LDL-R+ T cell could be utilized as a predictor for the proportion of TCR-T cells in the final cell product remained to be assessed.

In this study, we showed that LDL-R expression was significantly up-regulated with T cell activation by CD3/CD28-Dynabeads. The percentage of LDL-R+ T cells was directly proportional to that of the TCR-T cells post-activation. Also, we optimized the T cell activation method, through which a high proportion of TCR+ TCM cells after expansion could be achieved. Our findings suggest the possibility of LDL-R being an early evaluation parameter for the desired TCR-T cell products, contributing to an overall optimization strategy for the manufacturing process of adoptive therapy.

Materials and Methods

Cell lines and human PBMCs. Lenti-X 293T cells were purchased from Takara Biomedical Technology and cultured in DMEM (Gibco, Invitrogen, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Excell bio, Shanghai, PR China), 1 mM sodium pyruvate solution and 4 mM L-glutamine (Gibco, Invitrogen, CA, USA). The cells were then cultured at 37°C in a humidified incubator with 5 % CO2.

PBMC isolation. The whole blood was centrifuged for 15 mins at 400 g to separate the cellular fraction and plasma. The PBMCs were isolated via density gradient centrifugation with density gradient medium according to the manufacturer’s instructions (Lymphoprep, STEMCELL Technologies, Vancouver, Canada). Isolated PBMCs were cryopreserved and stored in liquid nitrogen until their use in the assays.

Lentivirus production. The gene sequences of mTCR, comprised of mouse alpha and beta TCR constant regions and human TCR variable regions, were cloned into the lentivirus vector pWPXL (Addgene Plasmid #12257). When grown to 80% confluency, Lenti-X293T cells were transfected with plasmids pWPXL, packaging plasmids psPAX2 (Addgene Plasmid #12260) and pMD2.G (Addgene Plasmid #12259) at a ratio of 5:2:1, using the Xfect Transfection Reagent (Takara, Kusatsu, Japan) following the manufacturer’s instructions. After culturing for 48 h, the supernatant was harvested and filtered by a 0.45 μm filter, then stored in −80°C for further analysis.

Activation of human PBMCs. The PBMCs were seeded in tissue culture-treated 24-well plates at density of 2×106 cells per well. CD3/CD28-Dynabeads (Thermo Fisher Scientific, Loughborough, UK) or anti-CD3 mAb (Miltenyi Biotec, Bergisch Gladbach, Germany, plate-bound, 1 μg/ml) and anti-CD28 mAb (Miltenyi Biotec, soluble, 1 μg/ml) were then added to the plates for different culturing periods and different doses, according to the experimental design. These cells were cultured in complete medium (90% RPMI-1640, Gibco, Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco, Invitrogen), 25 mM HEPES (Gibco, Invitrogen), 2 mM L-glutamine (Gibco, Invitrogen), 55 μM β-mercaptoethanol (Sigma, St. Louis, MO, USA), 100 μg/ml streptomycin (Gibco, Invitrogen), 100 U/ml penicillin (Gibco, Invitrogen), 10 ng/ml IL-7 (PreproTech, Rocky Hill, NJ, USA) and 10 ng/ml IL-15 (PreproTech). These cells were used for the transfection of mTCR and further expansion.

Transfection and expansion of human PBMCs. After activation, 2×105 stimulated PBMCs were seeded in 24-well tissue culture-treated plates. To these cells, 500 μl of lentiviral supernatant was added, containing mTCR gene after the lentivirus titer was measured and diluted to a common titer. The supernatant was replaced with fresh complement medium after 24 h. During the expansion, these cells were cultured at 37°C with 5% CO2 and cytokines (10 ng/ml IL-7 (PreproTech) and 10 ng/ml IL-15 (PreproTech) were added every other day. These cells were transferred to 6-well tissue culture plates depending on the total cell number. On the 5th and 14th day after transfection, these T cells were taken for subsequent experiments.

Flow cytometry analysis. 5×105 cells per tube were collected in PBS containing 2% FBS and stained with the anti-human antibody cocktail for 30 mins at room temperature in the dark. The following antibodies used for the T cell subsets marker assay were obtained from Biolegend, CA, USA: PE-anti-human CD3 antibody (Cat. 344806), BV510-anti-human CD3 antibody (Cat. 344828), Percy5.5-anti-human CD4 antibody (Cat. 300530), Alex700-anti-human CD8 antibody (Cat. 300920), FITC-anti-human CD8 antibody (Cat. 300906), BV421-anti-human CCR7 antibody (Cat. 353208), FITC-anti-human CD45RA antibody (Cat. 304106). The APC-anti-human LDL-R antibody (R&D Systems, Minneapolis, MN, USA, Cat. FAB2148A) was used for labeling activated T cells. The mouse TCRβ antibody was used for the identification of mTCR expression. Data was collected using a FACS Celesta flow cytometer (BD Biosciences, San Diego, CA, USA), and the results were analyzed using FlowJo software (FlowJo, LLC, BD Pharmingen, Franklin Lakes, NJ, USA). The T cell subtypes were analyzed for the following marker expression: Naive T cells (TN): CD45RA+CCR7+, TCM: CD45RA−CCR7+, Effector memory T cells (TEM): CD45RA−CCR7−, Effector T cells (TE): CD45RA+CCR7−.

Statistical analysis. GraphPad Prism 8.0 (GraphPad Software, San Diego, CA, USA) was used for statistical computations. Data are expressed as mean±standard error of the mean (SEM). Comparisons between groups were performed via the Student’s t-test for two groups, ANOVA and Tukey’s multiple comparisons post hoc test for three groups. Differences were considered statistically significant at p<0.05. *p<0.05, **p<0.01.

Results

The LDL-R+CD8+ T cell proportion was positively associated with the percentage of mTCR+CD8+ T cells post-transfection. To investigate the relationship between LDL-R expression and the efficiency of TCR transfection, we examined the proportion of LDL-R+ T cells and mTCR+ T cells in activated PBMCs via flow cytometry. The presence of LDL-R+ T cells was readily detectable post activation by OKT3 or CD3/CD28-Dynabeads, with the proportion of 67.7% and 60.8% in CD8+ T cell populations, respectively (Figure 1A). In CD4+ T cells, the proportion of LDL-R+ T cells was shown to be 50% when stimulated by OKT3 and 40% with Dynabeads activation (Figure 1A). No statistical significance was found in the LDL-R+ T cells proportions under these means of activation (Figure 1A).

Figure 1.
Figure 1.

The percentage of LDL-R+CD8+ T cells was directly proportional to that of mTCR+CD8+ T cells under CD3/CD28-Dynabeads activation. (A) The LDL-R expression on the membrane of CD4+ and CD8+ T cells was measured by flow cytometry post-activation by OKT3 or by CD3/CD28-Dynabeads for 48 h. (B) The proportions of mTCR+ T cells were assessed by flow cytometry three days post-transfection. Data are shown as mean±SEM of three independent healthy donors (n=3). Statistical significance was calculated using the Student t-test. *p<0.05 and **p<0.01.

Subsequently, we accessed the establishment of mTCR+ T cells post transfection. After a 3-day transfection period, the proportions of mTCR+ T cells were approximately 35% for both CD8+ and CD4+ T cells activated by OKT3 (Figure 1B). Comparatively, a significant elevation of mTCR+ T cell proportion was found in the group of cells activated by CD3/CD28-Dynabeads, resulting in 53.4% of mTCR+CD8+ T cells and 67.6% mTCR+CD4+ T cells (Figure 1B). Although comparable levels of LDL-R expression were observed in CD8+ and CD4+ T cells activated by these methods, the proportion of mTCR+ T cells in CD8+ T cells activated via CD3/CD28-Dynabeads was similarly high to that of the corresponding LDL-R+ T cells (Figure 1A and B). These findings suggested that activation via CD3/CD28-Dynabeads was a more efficient way to induce mTCR+ T cell products, and a high percentage of LDL-R+ T cells was directly proportional to that of mTCR+CD8+ T cells, but not mTCR+CD4+ T cells.

A 24 h-activation period was more effective to yield high proportion of mTCR+CD8+ T cells post transfection. Given that LDL-R expression was a rapid response post T cell activation, we examined the proportion of LDL-R+ T cells under a time course after the stimulation by CD3/CD28-Dynabeads. Flow cytometric results showed that the percentages of LDL-R+ T cells were robustly induced by 24 h, as high as 70.5% in CD8+ T cell populations and 59.1% in CD4+ T cells (Figure 2A). An additional activation period to 48 h did not alter the proportions of LDL-R+ T cells significantly (Figure 2A). Accordingly, we compared the effect of different activation periods on the proportions of mTCR-expressing T cells 3 days post-transfection. No apparent difference was observed in the mTCR+CD4+ T cells percentages under Dynabeads stimulation for 24 h and 48 h; however, an activation of 24 h induced more than 1.1-fold of the mTCR+CD8+ T cell proportion of the 48 h activation group (Figure 2B). These data suggest that CD3/CD28-Dynabeads stimulation for 24 h was sufficient to promote LDL-R expression in T cells and reducing the activation duration from 48 h to 24 h might be beneficial for the production of mTCR+CD8+ post-transfection.

Figure 2.
Figure 2.

An activation period of 24 h with CD3/CD28-Dynabeads was sufficient to induce LDL-R expression and mTCR+CD8+ T cell production. (A) The proportions of LDL-R+ T cells in CD4+ and CD8+ T cells were detected by flow cytometry after the stimulation of PBMCs for 24 and 48 h. Non-stimulated cells were used as control. (B) The percentages of mTCR+ T cells in CD4+ and CD8+ T cells were detected by flow cytometry under 24 and 48 h activation by CD3/CD28-Dynabeads. Data are shown as mean±SEM of three independent healthy donors (n=3). Statistical significance was calculated using ANOVA and Tukey’s multiple comparisons post hoc test in Figure 2A and Student t-test in Figure 2B. *p<0.05 and **p<0.01.

LDL-R+ T cell proportion was directly correlated with the percentage of mTCR+ T cells. As we had shown that the activation step played a vital role in determining the success establishment of mTCR+ T cell populations, we further verified whether the dosage of CD3/CD28-Dynabeads would be an influencing factor during this process. PBMCs were exposed to various ratios of CD3/CD28-Dynabeads; cells and the correlation between the proportion of LDL-R+ and mTCR+ T cells in CD8+ T cells were examined. Only 30% of LDL-R+ T were detected with a ratio of 0.5:1 (Figure 3A). When the ratio was increased to 1:1, the proportion of LDL-R+ T cells was significantly elevated in both CD8+ and CD4+ T cells, while continuing to increase the ratio to 2:1 did not further amplify the percentage of LDL-R-expressing T cells (Figure 3A).

Figure 3.
Figure 3.

The LDL-R+ T cell proportion might be predictive of the proportion of mTCR+ T cells under CD3/CD28-dynabeads activation. (A) The proportions of LDL-R+ T cells in CD4+ T cells and CD8+ T cells were detected by flow cytometry post-activation with different dosage ratios of CD3/CD28-Dynabeads : PBMCs for 24 h. (B) The percentage of mTCR+ T cells in CD3+ T cells was detected by flow cytometry after transfection under different activating conditions. Data were shown as mean±SEM of three independent healthy donors (n=3). (C) The correlation between the proportions of LDL-R+ T cells and mTCR+ T cells in CD8+ T cells was determined by linear regression. Bold indicate the data assessed from the optimized condition of the activation duration (24 h) and dosage (1:1). Data are shown as mean±SEM of three independent healthy donors (n=3). Statistical significance was calculated using ANOVA and Tukey’s multiple comparisons post hoc test in Figure 3A and 3B. Pearson’s correlation coefficient is used to identify the correlation in Figure 3C. *p<0.05 and **p<0.01.

Consistently, we found that the proportions of mTCR+CD8+ T cells post transfection were significantly higher under the activation conditions with a ratio of 1:1 than that of 0.5:1, and the 2:1 condition did not induce a profound enhancement of mTCR+CD8+ T cell proportion (Figure 3B). Meanwhile, no apparent changes in mTCR+CD4+ T cell proportions were observed among activating conditions with different Dynabeads:T cell ratios, despite the dose-dependent increase of LDL-R+ T cell proportions with Dynabeads (Figure 3B). Further, we compared the association of LDL-R+ T cell proportion and the post transfection mTCR+ T cell percentage, and found they exhibited a positive correlation only in the CD8+ T cell population, but not in the CD4+ T cells (Figure 3C). These results indicated that the percentage of LDL-R+ T cells after activation might be predictive of the post-transfection mTCR+ T cell proportion, facilitating the optimization of the stimulating process with CD3/CD28-dynabeads.

The proportions of mTCR+CD8+ TN cells were elevated with Dynabeads removal post-expansion. Stem-cell-like T cells possessing self-renewing and multipotent capabilities, such as TN and TCM, are desired components in TCR-T cell products (22). Therefore, we determined whether LDL-R+ T cell proportions could represent the proportions of mTCR+ TN and TCM cell populations. Flow cytometry analyses showed that the percentages of LDL-R+ T cells in TN, TCM, TEM and TE cell populations were all significantly enriched to over 50% after activation by CD3/CD28-Dynabeads for 24 h (Figure 4A). Extended activation time to 48 h did not affect the LDL-R+ T cell proportions in all but the TE cell population, which exhibited a 50% reduction (Figure 4A). Next, we subjected the activated cells to a rapid expansion period of 10 days to recapture the necessary step of pre-infusion TCR-T cell production. The proportions of mTCR+CD8+ T cells post expansion were determined and we found that the 24 h Dynabeads activation condition produced approximately 25% more mTCR+CD8+ T cells than the 48 h condition in the final product (Figure 4B). In contrast, the results showed that the additional 24 h in activation period produced more than 20% increase in the percentage of mTCR+CD4+ T cells post-expansion (Figure 4B). Analysis of the CD8+ T cell subtypes with 24 h activation demonstrated that the percentages of TCM cells in the final product were significantly higher than that of the TE cells (Figure 4C). Further, we found a positive correlation between the percentage of LDL-R+ T cells and that of CD8+ T cells (Figure 4D). These results suggest that the LDL-R+ T cell proportion might be indicative of the ratio of mTCR+ T cells in the expanded cell population, highlighting its possibility as an early assessment parameter for the TCR-T cell production process.

Figure 4.
Figure 4.
Figure 4.

The mTCR+CD8+ TN cell proportion was elevated with Dynabeads removal post-activation. (A) The LDL-R+ proportion of CD8+ T cell subtypes was measured by flow cytometry after activation with CD3/CD28-Dynabeads for 24 or 48 h. (B) mTCR+ T cells were transfected after activation by CD3/CD28-Dynabeads at a dose ratio of 1:1 for 24 or 48 h and expanded for 14 days. The proportion of CD4+ T cells and CD8+ T cells in mTCR+ T cells were determined by flow cytometry. (C) The mTCR+ CD8+ T cells percentage in the T cell subtypes was assessed by flow cytometry at day 10. (D) The correlation between the proportions of LDL-R+ T cells and mTCR+ T cells in CD8+ T cells and their subtypes was determined by linear regression. (E and F) After expansion for 10 days, the CD8+ T cell subtype composition (E) and the mTCR+CD8+ T cell percentage (F) were assessed in all subtypes with or without post-activation Dynabeads removal. Data are shown as SEM of three independent healthy donors (n=3). Statistical significance was calculated using one-way ANOVA and Tukey’s multiple comparisons post hoc test in Figure 4A and 4C and Student t-test in Figure 4E. Pearson’s correlation coefficient is used to identify the correlation in Figure 4D. *p<0.05 and **p<0.01.

Moreover, we tested whether the critical step of Dynabeads removal prior to rapid expansion could affect the T cell subtype composition in the final product. We observed a significant increase in the TN percentage and a significant reduction in the TE proportion with the inclusion of Dynabeads removal post-activation during production (Figure 4E). Also, we found that Dynabeads removal did not induce significant changes in the proportion of mTCR+ T cells in all subtypes, suggesting that the exclusion of Dynabeads might be beneficial in maintaining the high mTCR-expressing CD8+ TN cell population in the final product (Figure 4F). Collectively, these findings demonstrate that an optimized CD3/CD28-Dynabeads activation process could effectively increase the percentage of mTCR+CD8+ T cells in the final cell product, which could be predicted by the LDL-R+CD8+ T cell proportion as early as 3 days post-activation.

Discussion

The TCR-T cell quality in the final cell product is critical for its anti-tumor activity in vivo, as it is directly linked to the post infusion efficacy in tumor eradication (23, 24). For the multi-step process of TCR-T cell production, early evaluation parameters of the TCR-T proportion and phenotype would aid in controlling the quality and cost of the desired cell product. In this study, we established a positive correlation between the proportion of LDL-R+CD8+ T cells post-activation and the proportion of mTCR+CD8+ T cells in the final cell product, suggesting a possible new early evaluation parameter for the production of TCR-T cells applied in adoptive therapy.

The CD3/CD28 antibody-mediated activation step primes T cells for TCR introduction and cell expansion, and therefore serves as an essential step for TCR-T cell production (8, 11, 15, 16). We compared different conditions for T cell activation and generated an optimized strategy for this critical step with a better yield of the desired mTCR+CD8+ T cells. The use of cell-sized magnetic beads instead of plate-bound antibodies might facilitate the aggregation of activated T cells and induce high activation intensity (8, 15, 16). The shift of the classic 48 h activation period to 24 h was shown to induce sufficient elevation in LDL-R expression, which might be a prerequisite for subsequent TCR-delivery through viral transfection in CD8+ T cells. This was confirmed by the observed correlation of the LDL-R+CD8+ T cell proportion post-activation and the mTCR+CD8+ T cell proportion post-transfection. Mechanistically, it has been observed that LDL-R acts as the receptor of the lipoprotein, including cholesterol, which plays a role in regulating lysosomal response and mTORC1 activation in activated CD8+ T cells, but not in activated CD4+ T cells, and subsequently coordinates CD8+ T cell activation (19, 20). Conversely, the impact of LDL-R on the activation process of CD4+ T cells remains ambiguous, despite the identification of LDL’s ability to induce a central memory phenotype and a less exhausted phenotype in CD4+ T cells when added extrinsically in vitro during CD3/CD28 dynabeads mediated-activation (25). Besides, the uptake of glycolipid via the LDL-R and LDL receptor-related protein in antigen presenting cells leads to increased invariant natural killer T cell activation. This suggests that the transporter of lipid protein via LDL-R and the related protein may play a crucial role in the T cell activation, needing further investigation into the underlying mechanism (26).

Another important achievement with the optimized activation method was the selected enhancement of CD8+ TN and TCM proportions in the final product. It is likely that the relatively shorter stimulation period favors the maintenance of less differentiated cell populations prior to TCR gene transfection and cell expansion. These cells possess better potential to proliferate during the production process, and eventually, dominate the CD8+ T cell population. This is particularly important because the stemness of T cells expressing TCRs with high functional avidity, such as the one tested in this study, dictates their in vivo longevity and anti-tumor efficacy. Therefore, modifications of the T cell production process that could increase the desired TCM proportion would be practical and beneficial for the ultimate goal of adoptive therapy. Nevertheless, further investigation of the in vitro and in vivo cytotoxic functions of TCR-T cells established with our optimized method would be necessary.

Conclusion

In summary, our investigation found the proportion of LDL-R+CD8+ T cells post -activation was positively associated with the percentage of mTCR+CD8+ T cells post-transfection. The LDL-R+CD8+ T cells proportion served as an early assessment parameter for evaluating the desired TCR-expressing CD8+ T population in the final cell product for adoptive therapy. The method of T cell activation was optimized to achieve the T cell product with a less-differentiated phenotype. These findings offer crucial insights for the timely assessment of TCR-T quality and strategic optimizations of TCR-T cell production process.

Acknowledgements

We thank all healthy individuals who participated in this study. We thank Jingjing Huang for technical assistance and for secretarial work. This study was supported by the Chongqing Medical University fund (X4457) with the donation from Mr. Yuling Feng. This work was also supported by the National Natural Science Foundation of China (grant number 82271879) and the Chongqing Postdoctoral Science Foundation (cstc2020jcyj-bshX0060).

Footnotes

  • Authors’ Contributions

    Conceptualization, Y.W. (Yi Wang), Y.H., T.L. and A.J.; Data curation, Y.W. (Yi Wang) and Y.H.; Formal analysis, Y.W. (Yi Wang), Y.H., M.S., X.H., Y.W. (Yingming Wang), S.C. and J.W.; Funding acquisition, T.L. and A.J.; Investigation, M.S. and Y.W. (Yingming Wang); Methodology, Y.W. (Yi Wang), Y.H. and X.H.; Project administration, T.L.; Resources, A.J.; Supervision, A.J.; Validation, M.S.; Visualization, Y.W. (Yi Wang), Y.H., S.C. and J.W.; Writing – original draft, W.W. and T.L.; Writing – review & editing, W.W., T.L. and A.J.

  • Conflicts of Interest

    The Authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

  • Received July 13, 2023.
  • Revision received August 25, 2023.
  • Accepted August 29, 2023.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

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The Expression of LDL-R in CD8+ T Cells Serves as an Early Assessment Parameter for the Production of TCR-T Cells
YI WANG, YA-NAN HAO, MEI-YING SHEN, XIAO-JIAN HAN, YING-MING WANG, SI-YIN CHEN, JUN-FAN WANG, WANG WANG, TING-TING LI, AI-SHUN JIN
In Vivo Nov 2023, 37 (6) 2480-2489; DOI: 10.21873/invivo.13355
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