Identification of cross‐reactive antibodies for the detection of lymphocytes, myeloid cells and haematopoietic precursors in the naked mole rat

Abstract

The naked mole rat (Heterocephalus glaber, NMR) is a rodent with exceptional longevity, low rates of age-related diseases and spontaneous carcinogenesis. The NMR represents an attractive animal model in longevity and cancer research, but there are no NMRspecific antibodies available to study its immune system with respect to age-and cancerrelated questions. Substantial homology of major NMR immune cell markers with those of Guinea pig, human and, to a lesser extent, mouse and rat origin are implicated for the existence of immunological cross-reactivity. We identified 10 antibodies recognising eight immunophenotypic markers expressed on the NMR's T and B lymphocytes, macrophages/monocytes and putative haematopoietic precursors and used them for an immunophenotyping of leukocyte subsets of peripheral blood, spleen and bone marrow samples. Overall, we found that the leukocyte composition of NMR peripheral blood is comparable to that of mice. Notably, the frequency of cytotoxic T cells was found to be lower in the NMR compared to corresponding mouse tissues and human blood. Antibodies used in the present paper are available either commercially or from the scientific community and will provide new opportunities for the NMR as a model system in ageingand cancer-related research areas.Keywords: granulocytes r haematopoietic precursors r lymphocytes r myeloid cells r naked mole rat Additional supporting information may be found online in the Supporting Information section at the end of the article. years, which is five times longer than expected based on its body size, and exhibits neither significant senescence nor an age-related increase in mortality [1]. Moreover, spontaneous carcinogenesis was observed only recently in aged animals [2,3]. Several intrinsic protective cancer-related molecular mechanisms of the NMR were reported, including activity of the INK4 locus and mutation of the ERAS oncogene [4, 5], but they were not related to the NMR immune system.

Establishment and Clinical Application of a Method for Detecting T Lymphocyte Subsets by Cellular Immunochip Technology

“…Performance evaluations such as accuracy and precision have passed the CFDA approval and at the same time have been approved by the US Patent Office. Compared with flow cytometry, the instruments and detection reagents are expensive, require professionals and require high maintenance; 21 MRI detection and tracking technology are tedious to operate, temporarily not suitable for clinical use; 3 in situ MHC tetramer staining detection technology is not yet mature; 4 circulation cells Counting analysis (FTCA1) is complicated and sophisticated, and the development is more challenging; 5 ELISPOT technology mainly detects T cells secreted cytokines, and the absolute counting function is lacking; 7 the microfluidic chip platform cell affinity chromatography detection is heavily dependent on fluorescent labeling and imaging. 22 The operation of this project is simple, fast and easy to master.…”

“… 1 , 2 Therefore, the CD cell count has a guiding role in the early diagnosis and treatment of diseases with low cellular immune function and the judgment of the course of the disease. Other T cell detection methods such as MRI detection tracking, 3 In Situ MHC Tetramer Staining, 4 The Flow-Through cell Counting Assay (FTCA1), 5 the optofluidic ring resonator, 6 Enzyme-Linked Immunospot Assays 7 and many more.…”

Alternative Animal Models of Aging Research

“…Various, partly overlapping mechanisms were suggested to contribute to longevity and cancer resistance (Shepard and Kissil, 2020), e.g., over-expression of alpha-2 macroglobulin (Thieme et al, 2015), efficient DNA damage repair (MacRae et al, 2015), apoptosis and autophagy of damaged cells (Zhao et al, 2014(Zhao et al, , 2016Evdokimov et al, 2018), cytoprotective pathways (Lewis et al, 2015) and proteasome activity (Rodriguez et al, 2014). This list continues with translational fidelity (Azpurua et al, 2013), telomere and epigenome maintenance (Tan et al, 2017;Shekhidem et al, 2019), post-translational deimination (Pamenter et al, 2019), well-maintained splicing regulation (Lee et al, 2020), long non-coding RNAs (Jiang et al, 2016), and a strong innate immune response due to a highly developed myeloid compartment (Hilton et al, 2019;Shebzukhov et al, 2019). The surprisingly low incidence of cancer (Shepard and Kissil, 2020) was associated in particular with early contact inhibition (Seluanov et al, 2009).…”

Surprisingly long survival of premature conclusions about naked mole‐rat biology

“…Naked mole‐rats have a remarkably low incidence of typical age‐related diseases such as sarcopenia (O'Connor et al ., 2002; Stoll et al ., 2016), osteoporosis (Buffenstein et al ., 1994), and cancer (Buffenstein, 2008). Naked mole‐rats effectively control and shape their microbiome (Debebe et al ., 2016), probably due to a highly developed myeloid compartment of the immune system (Cheng et al ., 2017;Hilton et al ., 2019; Shebzukhov et al ., 2019) and expression of cathelicidin‐like protein with antibacterial activity (Cho et al ., 2018). Adult individuals are rarely found to be ill, both in the wild (Hill et al ., 1957) and under laboratory conditions (Buffenstein, 2008).…”

“…However, naked mole‐rats are susceptible to viral infections (Ross‐Gillespie et al ., 2007; Artwohl et al ., 2009) and intracellular parasites such as Leishmania donovani (Githure, Gardener, & Kinoti, 1988). This is in accordance with the observation that the immune system of naked mole‐rats has a relatively low proportion of lymphocytes (in particular B cells and cytotoxic T cells) and a complete absence of natural killer (NK) cells (Hilton et al ., 2019; Shebzukhov et al ., 2019).…”