Induction of alopecia areata in C3H/HeJ mice using cryopreserved lymphocytes

Abstract

Background: Alopecia areata (AA) is an autoimmune disease resulting in non-scarring hair loss. Animal models are useful means to identify candidates for therapeutic agents. The C3H/HeJ mouse AA model induced via transferring cultured lymphoid cells isolated from AA-affected mice is widely used for AA research. However, this conventional method requires the continuous breeding of AA mice. Objective: We aimed to establish a new method to generate AA model using the transfer of cryopreserved cells, which allows the rapid induction of a large number of AA mice when needed. Methods: We cryopreserved lymph node cells soon after isolation from AA-affected mice and injected thawed-cultured cells into recipient mice. H&E staining, immunohistochemical staining, quantitative real-time PCR and ELISA were conducted to identify pathological characteristics. Flow cytometry was performed to reveal the profile of transferred cells. Results: More than 90 % of recipient mice developed AA-like hair loss and showed inflammatory cell infiltration around anagen hair follicles, markedly increased mRNA expressions of interferon-g, CXCL11, and granzyme B, and elevated interferon-α protein levels in the skin compared with naïve mice. Higher percentages of effector memory T cells and dendritic cells in transferred cells resulted in a higher incidence of AA. Conclusion: This is the first report to establish a method for generating AA mice using cryopreserved lymphocytes. These AA mice have similar pathological characteristics to AA mice generated with the conventional method and AA patients. This convenient and reproducible method is expected to be valuable for AA study.

Construction of regulatory network for alopecia areata progression and identification of immune monitoring genes based on multiple machine-learning algorithms

“…Hashimoto et al . 40 further showed that effector memory CD4 + T cells were able to stimulate native and CD8 + cells through antigen-presenting cells in axillary lymph nodes and direct T cells to attack hair follicles, leading to AA. In addition, although a significant decrease in neutrophils was observed in the severe AA group in this study, previous studies have shown that the ratio of neutrophils to lymphocytes was not a good indicator for evaluating the severity of AA.…”

Genome-Wide Association Study of Alopecia Areata in Taiwan: The Conflict Between Individuals and Hair Follicles

“… 31 There is a correlation between the gene variation site associated with the C3H-HeJ alopecia pattern in mice and the gene variation site of human leukocyte antigens (HLA). 32 Collectively, this evidence suggests that AA is inherited and may run in families. The genetic basis underlying AA in Taiwan has not yet been definitively examined.…”

Inhibition of T-cell activity in alopecia areata: recent developments and new directions

“…Tissue resident memory T cells have been hypothesized to be involved in AA pathogenesis and lesion recurrence (65). This notion is based on increased levels of these cells in AA lesions observed in some studies (65), including evidence from AA mouse models linking injection of high proportions of effector memory CD8+ T cells with increased incidence of AA development (66). More recently, a CD44 superhigh Cd49d low subset of CD8 + virtual memory T (T vm ) cells have been identified in stomach-draining lymph nodes of alopecic mice with the ability to promote disease upon transfer to naive recipient mice.…”