Generation of competent bone marrow-derived antigen presenting cells from the deer mouse (Peromyscus maniculatus).
BACKGROUND
Human infections with Sin Nombre virus (SNV) and related New World hantaviruses often lead to hantavirus cardiopulmonary syndrome (HCPS), a sometimes fatal illness. Lungs of patients who die from HCPS exhibit cytokine-producing mononuclear infiltrates and pronounced pulmonary inflammation. Deer mice (Peromyscus maniculatus) are the principal natural hosts of SNV, in which the virus establishes life-long persistence without conspicuous pathology. Little is known about the mechanisms SNV employs to evade the immune response of deer mice, and experimental examination of this question has been difficult because of a lack of methodologies for examining such responses during infection. One such deficiency is our inability to characterize T cell responses because susceptible syngeneic deer mice are not available.


RESULTS
To solve this problem, we have developed an in vitro method of expanding and generating competent antigen presenting cells (APC) from deer mouse bone marrow using commercially-available house mouse (Mus musculus) granulocyte-macrophage colony stimulating factor. These cells are capable of processing and presenting soluble protein to antigen-specific autologous helper T cells in vitro. Inclusion of antigen-specific deer mouse antibody augments T cell stimulation, presumably through Fc receptor-mediated endocytosis.


CONCLUSIONS
The use of these APC has allowed us to dramatically expand deer mouse helper T cells in culture and should permit extensive characterization of T cell epitopes. Considering the evolutionary divergence between deer mice and house mice, it is probable that this method will be useful to other investigators using unconventional models of rodent-borne diseases.
Davenport
Bennett J
BJ
Department of Biological Sciences, Mesa State College, 1100 North Ave., Grand Junction, CO 81501, USA. bennettdavenport@yahoo.com
Willis
Derall G
DG
Prescott
Joseph
J
Farrell
Regina M
RM
Coons
Teresa A
TA
Schountz
Tony
T
D000818:Animals; D017951:Antigen Presentation / Q000502:physiology; D000938:Antigen-Presenting Cells / Q000166:cytology / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D003001:Cloning, Molecular / Q000379:methods; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis; D006801:Humans; D007106:Immune Sera / Q000096:biosynthesis; D007376:Interleukin-2 / Q000276:immunology; D008213:Lymphocyte Activation / Q000502:physiology; D051379:Mice; D010542:Peromyscus / Q000235:genetics* / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis; D016176:T-Lymphocyte Subsets / Q000502:physiology; D006377:T-Lymphocytes, Helper-Inducer / Q000502:physiology
D018984:Epitopes, T-Lymphocyte; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D007106:Immune Sera; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin