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huPBMC humanized immunocompetent mouse modeling platform: simulates human immune responses and enables evaluation of the efficacy of immunotherapeutics.

　　Peripheral blood mononuclear cells (PBMCs) are nucleated cells circulating in the peripheral blood, primarily comprising lymphocytes (T cells or B cells), monocytes, phagocytes, dendritic cells, and a small number of other cell types. They constitute an essential cellular component of the body’s immune response.

　　Humanized mouse models of the immune system are generated by engrafting human hematopoietic cells, lymphocytes, or tissues into immunodeficient mice, thereby reconstituting a human immune system within the host. These models can effectively recapitulate human immune responses and more accurately mimic human immune characteristics, enabling the evaluation of the efficacy of anti‑tumor agents and facilitating the identification of promising therapeutics, thus offering hope for curing more cancer patients. They also provide a powerful platform for investigating the intricate interplay between human tumors and the immune system.

　　Modeling Protocol

　　| Experimental animal: Severe immunodeficient mice—NAG

　　| Experimental Protocol:

　　① After determining the appropriate cell injection dose, thaw the cells according to the supplier’s instructions and adjust the concentration prior to cell injection.

　　② Place the cells in a 4°C ice pack and transport them to the facility for injection (tail vein injection or intraperitoneal injection); after inoculation, monitor the mice’s condition.

　　| Experimental Evaluation System:

　　01. GvHD Observation

　　Following inoculation of huPBMCs, GvHD was monitored twice weekly, and clinical scores were assigned based on observational assessments of body weight, mouse posture, activity level, fur condition, skin appearance, and other parameters.

　　

 

　　02. Flow Cytometry Flow Cytometry

　　Following vaccination, flow cytometry was performed weekly, with blood collected from the orbital sinus of mice. Flow cytometry was used to determine the proportions of mCD45‑positive cells as well as hCD45‑, hCD3‑, hCD4‑, and hCD8‑positive cells. The day of inoculation was designated Day 0, and samples were analyzed on Days 7, 14, 21, 28, and 35.

　　03. Survival Curve

　　Record the mortality of mice and plot a survival curve.

　　| Experimental Results:

　　

 

　　

 

　　Image source: Ailinfei

　　| This study evaluates the reconstruction efficacy of the huPBMC-NAG model 01

　　Based on statistical analysis of mouse survival at the experimental endpoint (day 29), huPBMC-donor1 exhibited the longest survival time in NAG mice.

　　02. Based on the analysis of GvHD scores, huPBMC-donor1 and huPBMC-donor2 elicited relatively severe GvHD responses in NAG mice.

　　03. Based on the analysis of peripheral blood immune cell proportions in D29-day-old mice, huPBMC-donor1 exhibited a higher engraftment rate in NAG mice.

　　| Advantages and Limitations of the huPBMC Humanized Mouse Model

　　Mouse models transduced with huPBMCs offer numerous advantages, making them widely used, including:

　　01. Mice transduced with huPBMCs constitute a T-cell–dependent model, and many immunotherapies involve T-cell modulation or re‑targeting.

　　The huPBMC humanized mouse model is relatively easy to establish and does not require specialized training or capital equipment.

　　02. The transplantation process is very fast.

　　The same immune cell donor can be used multiple times to facilitate longitudinal studies.

　　03. The mouse model transduced with human PBMCs is more cost-effective than other humanized models.

　　Despite the model’s popularity, there remains some hesitation about its use. This situation is primarily attributable to two factors:

　　① GvT is highly likely to inhibit tumor growth in this model. ② The occurrence of GvHD in this model shortened the duration of the study.

　　These two issues can, to some extent, be mitigated by selecting appropriate huPBMC donors, as the severity of both effects depends on the donor. Donor selection to mitigate GvT can be assessed through short-term co-culture assays, whereas donor selection to reduce GvHD requires in vivo experiments using xenograft models relevant to the tumor of interest.

　　Conclusion

　　The humanized mouse model based on huPBMCs involves transplanting human peripheral blood mononuclear cells (PBMCs) into severely immunocompromised mice, thereby mimicking the human immune system. Subsequently, an appropriate model is selected to reconstruct a human‑like immune microenvironment as closely as possible, enabling the simulation of immune responses and the progression of disease in vivo. Aniphe BioLab can, according to experimental requirements, select a suitable huPBMC‑based humanized model and conduct subsequent animal studies; its flexible experimental design allows it to provide customers with high‑quality models.

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