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Disease Models | Humanized Mouse Models with Human PBMCs in Immunotherapy

Peripheral blood mononuclear cell (PBMC) Namely, the mononuclear cells in peripheral blood, which are primarily composed of lymphocytes (T or B cells), monocytes, phagocytes, dendritic cells, and a few other cell types, 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 in 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 effective 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

| Laboratory animals: Severely immunocompromised 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:

0 1 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

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, hCD45‑positive, hCD3‑positive, hCD4‑positive, and hCD8‑positive cells. The day of inoculation was designated as Day 0, and samples were analyzed on Days 7, 14, 21, 28, and 35.

0 3 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.

0 1 Based on statistical analysis of mouse survival at the experimental endpoint (day 29), huPBMC-donor1 exhibited the longest survival time in NAG mice.

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

0 3 According to the analysis of peripheral blood immune cell proportions in D29-day-old mice, huPBMC-donor1 exhibited a favorable 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:

0 1 Mouse models transduced with human PBMCs are T-cell–dependent, and many immunotherapies involve T-cell modulation or re‑targeting.

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

0 3 The transplant time is very fast.

0 4 The same immune cell donor can be used multiple times to facilitate longitudinal studies.

0 5 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 In this model, the occurrence of GvHD 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 primarily 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 immune microenvironment as closely as possible, enabling the simulation of immune responses and the progression of diseases 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 clients with high‑quality models.

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