Leonard Shultz


  • PhD, University of Massachusetts.1972


Spontaneous mutations that cause defects in immunological function in mice have provided important research tools with which to investigate the development and regulation of the immune system. A number of these mutations serve as models for specific human diseases and provide critical tools to increase the understanding of human immunodeficiency diseases, autoimmunity and leukemia. Spontaneous as well as targeted immunological mutations also provide mouse models that support engraftment with human stem cells and with human peripheral blood lymphocytes. These “humanized” mice have provided critical models for experimental investigation of human immune diseases without putting individuals at risk. Our recent research has leveraged these mouse models for studies on diabetes, bone development, autoimmunity, malignancy, anemia, thrombocytopenia, infectious diseases, transplantation tolerance and asthma.

Research Interests

Development of Humanized SCID Mice and Studies of Single Gene Models for Human Hematological Diseases
Advances in our understanding of the development and regulation of the immune system in normal and pathologic states rely heavily on studies of mice with genetically determined immunodeficiency and autoimmunity. Research in our laboratory encompasses (1) basic studies focused on elucidating the mechanisms underlying immunodeficiency, autoimmunity, and hematological diseases; and (2) applied studies focused on the development of effective immunodeficient mouse models that support heightened engraftment with human hematopoietic stem cells. These “humanized SCID mice” will facilitate studies of human immunity, regenerative medicine, and neoplasia. Examples of the studies carried out over the last year are summarized below:

Development of Humanized Mice for Translational Biomedical Research
Complex biological processes often require in vivo analysis, and important research advances have been obtained using mice as a model system for the study of many biological systems. However, mice are not humans, and the study of human biology in vivo is severely limited by ethical and technical constraints. There is a growing need for animal models to carry out in vivo studies on human cells, tissues, and organs without putting individuals at risk. Humanized mice, or mouse–human chimeras, have been developed to overcome these limitations and have become an important research tool for the in vivo study of human cells and tissues. We have developed a NOD-Prkdcscid (NOD/SCID) mouse strain harboring a null mutation of the common cytokine receptor γ chain (Il2rgtm1Wjl) (NOD/SCID/IL2rγnull) (full name NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ). In collaboration with Dale Greiner and his colleagues at The University of Massachusetts Medical School along with Fumihiko Ishikawa and his colleagues at The Riken Research Center for Allergy and Immunology, we have shown that these mice can efficiently support development of all myelo-erythroid components and functional lymphoid cells following human hematopoietic stem cell (HSC) engraftment. Development of mice that are ‘humanized’ by engraftment of human tissues, HSCs, and peripheral-blood mononuclear cells (PBMCs) provides an opportunity to study human biological processes in vivo that would otherwise not be possible. The new generations of humanized mice are proving to be powerful tools in preclinical testing and in the investigation of many human biological processes. The culmination of decades of research on humanized mice is leading to advances in our understanding of human hematopoiesis, innate and adaptive immunity, autoimmunity, infectious diseases, cancer biology, and regenerative medicine. The potential for new advances in our understanding of human biological systems provided by studies in humanized mice remains promising. The development of humanized mice based on immunodeficient IL2rγnullhosts has overcome many of the limitations and constraints of previously available models.

The Mouse Mutation “Thrombocytopenia and Cardiomyopathy” (trac) is within the Abcg5 Gene: A Single Gene Model for Human Mediterranean Macrothrombocytopenia
Homozygosity for a new spontaneous mouse mutation named “thrombocytopenia and cardiomyopathy” (trac) results in thrombocytopenia, dilated cardiomyopathy, and infertility. A/J-trac/trac mice show a precipitous drop in platelet numbers and increases in platelet volume by 4 weeks of age. By 2-3 months of age, trac/trac mice have a 20-fold decrease in platelet number, a 3-fold increase in platelet volume, and a greatly increased bleeding time. Blood smears showed abnormally large platelets and megakaryocytoid cells. The trac/trac mice also developed mild microcytic anemia accompanied by the presence of stomatocytes, a doubling of reticulocyte numbers, and a two-fold decrease in WBC counts. Increased numbers of megakaryocytes were present in bone marrow, spleen, and lungs. Ultrastructural studies of trac/trac megakaryocytes showed a poorly developed demarcation system and a failure to form platelet territories. Thetrac/trac platelets were enlarged, spherical, and contained numerous small alpha granules. The thrombocytopenia was not associated with defects intrinsic to bone marrow progenitor cells. Although thrombopoietin (TPO) levels were decreased, TPO treatment failed to reverse the thrombocytopenia. To identify the responsible gene, we produced a fine-structure genetic map of a 5-megabase interval containing the trac locus on mouse chromosome 17. The human syntenic region is Chr 2p21-p22. Analyses of 1100 F2 progeny from intercross matings of (A/J x C57BL/6J) F1 +/trac mice narrowed the interval to 0.3 Mb containing 4 genes. Sequencing of these genes revealed a G-to-A mutation at base 1435 (refseq nm031884) of Abcg5 (ATP-binding cassette sub-family G, member 5). This G>A base change results in a tryptophan codon (UGG) at amino acid position 463(uniprot) being changed to a premature stop codon (UAG). The transmembrane helices prediction program, TMHMM, predicts that the premature stop codon would truncate the last four of the six transmembrane domains of the ABCG5 protein. No DNA alterations were found in any of the other candidate genes. Genetic crosses of +/trac mice with mice doubly transgenic for the closely linked human ABCG5 and ABCG8 genes (stock B6SJL-Tg(ABCG5/ABCG8)14-2Hobb/J) showed that the transgenes normalized platelet counts and volumes in trac/trac mice. ABCG5 (sterolin-1) functions as part of a heterodimer, with ABCG8, that regulates plant sterol uptake. Thetrac/trac mutant mice have greatly elevated plasma levels of plant sterols. When placed on a phytosterol-free diet, the thrombocytopenia was reversed. Recent studies have shown that Mediterranean Macrothrombocytopenia is caused by mutations in ABCG5 or ABCG8. Identification of the molecular basis of the mouse Abcg5trac mutation provides a new model for studying the role of phytosterols in pathogenic changes in the hematopoietic, cardiovascular, and reproductive systems.

Selected Publications

  • Bankert RB, Balu-Iyer SV, Odunsi K, Shultz LD, Kelleher RJ, Jr., Barnas JL, Simpson-Abelson M, Parsons R, Yokota SJ. 2011. Humanized mouse model of ovarian cancer recapitulates patient solid tumor progression, ascites formation, and metastasis. PLoS One 6: e24420
  • Dash Y, Ramesh M, Kalyanasundaram R, Munirathinam G, Shultz LD, Rajan TV. 2011. Granuloma formation around filarial larvae triggered by host responses to an excretory/secretory antigen. Infect Immun 79: 838-845
  • Diiorio P, Jurczyk A, Yang C, Racki WJ, Brehm MA, Atkinson MA, Powers AC, Shultz LD, Greiner DL, Bortell R. 2011. Hyperglycemia-Induced Proliferation of Adult Human Beta Cells Engrafted Into Spontaneously Diabetic Immunodeficient NOD-Rag1null IL2rgammanull Ins2Akita Mice. Pancreas 40: 1147-1149
  • Dorrell C, Grompe MT, Pan FC, Zhong Y, Canaday PS, Shultz LD, Greiner DL, Wright CV, Streeter PR, Grompe M. 2011. Isolation of mouse pancreatic alpha, beta, duct and acinar populations with cell surface markers. Mol Cell Endocrinol 339: 144-150
  • Foreman O, Kavirayani AM, Griffey SM, Reader R, Shultz LD. 2011. Opportunistic bacterial infections in breeding colonies of the NSG mouse strain. Vet Pathol 48: 495-499
  • Liu G, Dou S, Cheng D, Leif J, Rusckowski M, Streeter PR, Shultz LD, Hnatowich DJ, Greiner DL. 2011. Human islet cell MORF/cMORF pretargeting in a xenogeneic murine transplant model. Mol Pharm 8: 767-773
  • Schleifman EB, Bindra R, Leif J, Del Campo J, Rogers FA, Uchil P, Kutsch O, Shultz LD, Kumar P, Greiner DL, Glazer PM. 2011. Targeted Disruption of the CCR5 Gene in Human Hematopoietic Stem Cells Stimulated by Peptide Nucleic Acids. Chem Biol 18: 1189-1198
  • Whitfield-Larry F, Young EF, Talmage G, Fudge E, Azam A, Patel S, Largay J, Byrd W, Buse J, Calikoglu AS, Shultz LD, Frelinger JA. 2011. HLA-A2-matched peripheral blood mononuclear cells from type 1 diabetic patients, but not nondiabetic donors, transfer insulitis to NOD-scid/gammac(null)/HLA-A2 transgenic mice concurrent with the expansion of islet-specific CD8+ T cells. Diabetes 60: 1726-1733
  • Zhao L, Spassieva SD, Jucius TJ, Shultz LD, Shick HE, Macklin WB, Hannun YA, Obeid LM, Ackerman SL. 2011. A deficiency of ceramide biosynthesis causes cerebellar purkinje cell neurodegeneration and lipofuscin accumulation. PLoS Genet 7: e1002063
  • Brehm MA, Bortell R, Diiorio P, Leif J, Laning J, Cuthbert A, Yang C, Herlihy M, Burzenski L, Gott B, Foreman O, Powers AC, Greiner DL, Shultz LD. 2010. Human immune system development and rejection of human islet allografts in spontaneously diabetic NOD-Rag1null IL2rgammanull Ins2Akita mice. Diabetes 59: 2265-2270
  • Brehm MA, Cuthbert A, Yang C, Miller DM, DiIorio P, Laning J, Burzenski L, Gott B, Foreman O, Kavirayani A, Herlihy M, Rossini AA, Shultz LD, Greiner DL. 2010. Parameters for establishing humanized mouse models to study human immunity: analysis of human hematopoietic stem cell engraftment in three immunodeficient strains of mice bearing the IL2rgamma(null) mutation. Clin Immunol 135: 84-98
  • Brehm MA, Shultz LD, Greiner DL. 2010. Humanized mouse models to study human diseases. Curr Opin Endocrinol Diabetes Obes 17: 120-125
  • Chase TH, Lyons BL, Bronson RT, Foreman O, Donahue LR, Burzenski LM, Gott B, Lane P, Harris B, Ceglarek U, Thiery J, Wittenburg H, Thon JN, Italiano JE, Jr., Johnson KR, Shultz LD. 2010. The mouse mutation “thrombocytopenia and cardiomyopathy” (trac) disrupts Abcg5: a spontaneous single gene model for human hereditary phytosterolemia/sitosterolemia. Blood 115: 1267-1276
  • Gori JL, Tian X, Swanson D, Gunther R, Shultz LD, McIvor RS, Kaufman DS. 2010. In vivo selection of human embryonic stem cell-derived cells expressing methotrexate-resistant dihydrofolate reductase. Gene Ther 17: 238-249
  • Libby SJ, Brehm MA, Greiner DL, Shultz LD, McClelland M, Smith KD, Cookson BT, Karlinsey JE, Kinkel TL, Porwollik S, Canals R, Cummings LA, Fang FC. 2010. Humanized nonobese diabetic-scid IL2rgammanull mice are susceptible to lethal Salmonella Typhi infection. Proc Natl Acad Sci U S A 107: 15589-15594
  • Moriceau G, Ory B, Mitrofan L, Riganti C, Blanchard F, Brion R, Charrier C, Battaglia S, Pilet P, Denis MG, Shultz LD, Monkkonen J, Redini F, Heymann D. 2010. Zoledronic acid potentiates mTOR inhibition and abolishes the resistance of osteosarcoma cells to RAD001 (Everolimus): pivotal role of the prenylation process. Cancer Res 70: 10329-10339
  • Morisot S, Wayne AS, Bohana-Kashtan O, Kaplan IM, Gocke CD, Hildreth R, Stetler-Stevenson M, Walker RL, Davis S, Meltzer PS, Wheelan SJ, Brown P, Jones RJ, Shultz LD, Civin CI. 2010. High frequencies of leukemia stem cells in poor-outcome childhood precursor-B acute lymphoblastic leukemias. Leukemia 24: 1859-1866
  • Pino S, Brehm MA, Covassin-Barberis L, King M, Gott B, Chase TH, Wagner J, Burzenski L, Foreman O, Greiner DL, Shultz LD. 2010. Development of novel major histocompatibility complex class I and class II-deficient NOD-SCID IL2R gamma chain knockout mice for modeling human xenogeneic graft-versus-host disease. Methods Mol Biol 602: 105-117
  • Racki WJ, Covassin L, Brehm M, Pino S, Ignotz R, Dunn R, Laning J, Graves SK, Rossini AA, Shultz LD, Greiner DL. 2010. NOD-scid IL2rgamma(null) mouse model of human skin transplantation and allograft rejection. Transplantation 89: 527-536
  • Saito Y, Kitamura H, Hijikata A, Tomizawa-Murasawa M, Tanaka S, Takagi S, Uchida N, Suzuki N, Sone A, Najima Y, Ozawa H, Wake A, Taniguchi S, Shultz LD, Ohara O, Ishikawa F. 2010. Identification of therapeutic targets for quiescent, chemotherapy-resistant human leukemia stem cells. Sci Transl Med 2: 17ra19
  • Saito Y, Uchida N, Tanaka S, Suzuki N, Tomizawa-Murasawa M, Sone A, Najima Y, Takagi S, Aoki Y, Wake A, Taniguchi S, Shultz LD, Ishikawa F. 2010. Induction of cell cycle entry eliminates human leukemia stem cells in a mouse model of AML. Nat Biotechnol 28: 275-280
  • Shultz LD, Saito Y, Najima Y, Tanaka S, Ochi T, Tomizawa M, Doi T, Sone A, Suzuki N, Fujiwara H, Yasukawa M, Ishikawa F. 2010. Generation of functional human T-cell subsets with HLA-restricted immune responses in HLA class I expressing NOD/SCID/IL2r gamma(null) humanized mice. Proc Natl Acad Sci U S A 107: 13022-13027
  • Subramanya S, Kim SS, Abraham S, Yao J, Kumar M, Kumar P, Haridas V, Lee SK, Shultz LD, Greiner D, N M, Shankar P. 2010. Targeted delivery of small interfering RNA to human dendritic cells to suppress dengue virus infection and associated proinflammatory cytokine production. J Virol 84: 2490-2501
  • Zhang B, Strauss AC, Chu S, Li M, Ho Y, Shiang KD, Snyder DS, Huettner CS, Shultz L, Holyoake T, Bhatia R. 2010. Effective targeting of quiescent chronic myelogenous leukemia stem cells by histone deacetylase inhibitors in combination with imatinib mesylate. Cancer Cell 17: 427-442
  • Bernstein JM, Brooks SP, Lehman HK, Pope L, Sands A, Shultz LD, Bankert RB. 2009. Human nasal polyp microenvironments maintained in a viable and functional state as xenografts in NOD-scid IL2rgamma(null) mice. Ann Otol Rhinol Laryngol 118: 866-875
  • Chase TH, Cox GA, Burzenski L, Foreman O, Shultz LD. 2009. Dysferlin deficiency and the development of cardiomyopathy in a mouse model of limb-girdle muscular dystrophy 2B. Am J Pathol 175: 2299-2308
  • Chopra P, Diiorio P, Pino SC, Wilson SB, Phillips NE, Mordes JP, Rossini AA, Greiner DL, Shultz LD, Bortell R. 2009. Failure of alpha-galactosylceramide to prevent diabetes in virus-inducible models of type 1 diabetes in the rat. In Vivo 23: 195-201
  • Jaiswal S, Pearson T, Friberg H, Shultz LD, Greiner DL, Rothman AL, Mathew A. 2009. Dengue virus infection and virus-specific HLA-A2 restricted immune responses in humanized NOD-scid IL2rgammanull mice. PLoS One 4: e7251
  • Jimenez-Diaz MB, Mulet T, Viera S, Gomez V, Garuti H, Ibanez J, Alvarez-Doval A, Shultz LD, Martinez A, Gargallo-Viola D, Angulo-Barturen I. 2009. Improved murine model of malaria using Plasmodium falciparum competent strains and non-myelodepleted NOD-scid IL2Rgammanull mice engrafted with human erythrocytes. Antimicrob Agents Chemother 53: 4533-4536
  • King MA, Covassin L, Brehm MA, Racki W, Pearson T, Leif J, Laning J, Fodor W, Foreman O, Burzenski L, Chase TH, Gott B, Rossini AA, Bortell R, Shultz LD, Greiner DL. 2009. Human peripheral blood leucocyte non-obese diabetic-severe combined immunodeficiency interleukin-2 receptor gamma chain gene mouse model of xenogeneic graft-versus-host-like disease and the role of host major histocompatibility complex. Clin Exp Immunol 157: 104-118
  • Mangada J, Pearson T, Brehm MA, Wicker LS, Peterson LB, Shultz LD, Serreze DV, Rossini AA, Greiner DL. 2009. Idd loci synergize to prolong islet allograft survival induced by costimulation blockade in NOD mice. Diabetes 58: 165-173
  • Miller DM, Thornley TB, Pearson T, Kruger AJ, Yamazaki M, Shultz LD, Welsh RM, Brehm MA, Rossini AA, Greiner DL. 2009. TLR agonists prevent the establishment of allogeneic hematopoietic chimerism in mice treated with costimulation blockade. J Immunol 182: 5547-5559
  • Tian X, Hexum MK, Penchev VR, Taylor RJ, Shultz LD, Kaufman DS. 2009. Bioluminescent imaging demonstrates that transplanted human embryonic stem cell-derived CD34(+) cells preferentially develop into endothelial cells. Stem Cells 27: 2675-2685
  • Unsinger J, McDonough JS, Shultz LD, Ferguson TA, Hotchkiss RS. 2009. Sepsis-induced human lymphocyte apoptosis and cytokine production in “humanized” mice. J Leukoc Biol 86: 219-227
  • Agliano A, Martin-Padura I, Mancuso P, Marighetti P, Rabascio C, Pruneri G, Shultz LD, Bertolini F. 2008. Human acute leukemia cells injected in NOD/LtSz-scid/IL-2Rgamma null mice generate a faster and more efficient disease compared to other NOD/scid-related strains. Int J Cancer 123: 2222-2227
  • Croker BA, Lawson BR, Rutschmann S, Berger M, Eidenschenk C, Blasius AL, Moresco EM, Sovath S, Cengia L, Shultz LD, Theofilopoulos AN, Pettersson S, Beutler BA. 2008. Inflammation and autoimmunity caused by a SHP1 mutation depend on IL-1, MyD88, and a microbial trigger. Proc Natl Acad Sci U S A 105: 15028-15033
  • Gaines P, Tien CW, Olins AL, Olins DE, Shultz LD, Carney L, Berliner N. 2008. Mouse neutrophils lacking lamin B-receptor expression exhibit aberrant development and lack critical functional responses. Exp Hematol 36: 965-976
  • Giassi LJ, Pearson T, Shultz LD, Laning J, Biber K, Kraus M, Woda BA, Schmidt MR, Woodland RT, Rossini AA, Greiner DL. 2008. Expanded CD34+ human umbilical cord blood cells generate multiple lymphohematopoietic lineages in NOD-scid IL2rgamma(null) mice. Exp Biol Med (Maywood) 233: 997-1012
  • Harris MA, Yang H, Low BE, Mukherjee J, Guha A, Bronson RT, Shultz LD, Israel MA, Yun K. 2008. Cancer stem cells are enriched in the side population cells in a mouse model of glioma. Cancer Res 68: 10051-10059
  • Ishikawa F, Saito Y, Yoshida S, Harada M, Shultz LD. 2008. The differentiative and regenerative properties of human hematopoietic stem/progenitor cells in NOD-SCID/IL2rgamma(null) mice. Curr Top Microbiol Immunol 324: 87-94
  • King M, Pearson T, Rossini AA, Shultz LD, Greiner DL. 2008. Humanized mice for the study of type 1 diabetes and beta cell function. Ann N Y Acad Sci 1150: 46-53
  • King M, Pearson T, Shultz LD, Leif J, Bottino R, Trucco M, Atkinson MA, Wasserfall C, Herold KC, Woodland RT, Schmidt MR, Woda BA, Thompson MJ, Rossini AA, Greiner DL. 2008. A new Hu-PBL model for the study of human islet alloreactivity based on NOD-scid mice bearing a targeted mutation in the IL-2 receptor gamma chain gene. Clin Immunol 126: 303-314
  • Kong Y, Yoshida S, Saito Y, Doi T, Nagatoshi Y, Fukata M, Saito N, Yang SM, Iwamoto C, Okamura J, Liu KY, Huang XJ, Lu DP, Shultz LD, Harada M, Ishikawa F. 2008. CD34+CD38+CD19+ as well as CD34+CD38-CD19+ cells are leukemia-initiating cells with self-renewal capacity in human B-precursor ALL. Leukemia 22: 1207-1213
  • Kumar P, Ban HS, Kim SS, Wu H, Pearson T, Greiner DL, Laouar A, Yao J, Haridas V, Habiro K, Yang YG, Jeong JH, Lee KY, Kim YH, Kim SW, Peipp M, Fey GH, Manjunath N, Shultz LD, Lee SK, Shankar P. 2008. T cell-specific siRNA delivery suppresses HIV-1 infection in humanized mice. Cell 134: 577-586
  • le Viseur C, Hotfilder M, Bomken S, Wilson K, Rottgers S, Schrauder A, Rosemann A, Irving J, Stam RW, Shultz LD, Harbott J, Jurgens H, Schrappe M, Pieters R, Vormoor J. 2008. In childhood acute lymphoblastic leukemia, blasts at different stages of immunophenotypic maturation have stem cell properties. Cancer Cell 14: 47-58
  • Mori K, Blanchard F, Charrier C, Battaglia S, Ando K, Duplomb L, Shultz LD, Redini F, Heymann D. 2008. Conditioned media from mouse osteosarcoma cells promote MC3T3-E1 cell proliferation using JAKs and PI3-K/Akt signal crosstalk. Cancer Sci 99: 2170-2176
  • Pearson T, Greiner DL, Shultz LD. 2008. Creation of “humanized” mice to study human immunity. Curr Protoc Immunol Chapter 15: Unit 15 21
  • Pearson T, Greiner DL, Shultz LD. 2008. Humanized SCID mouse models for biomedical research. Curr Top Microbiol Immunol 324: 25-51
  • Pearson T, Shultz LD, Lief J, Burzenski L, Gott B, Chase T, Foreman O, Rossini AA, Bottino R, Trucco M, Greiner DL. 2008. A new immunodeficient hyperglycaemic mouse model based on the Ins2Akita mutation for analyses of human islet and beta stem and progenitor cell function. Diabetologia 51: 1449-1456
  • Pearson T, Shultz LD, Miller D, King M, Laning J, Fodor W, Cuthbert A, Burzenski L, Gott B, Lyons B, Foreman O, Rossini AA, Greiner DL. 2008. Non-obese diabetic-recombination activating gene-1 (NOD-Rag1 null) interleukin (IL)-2 receptor common gamma chain (IL2r gamma null) null mice: a radioresistant model for human lymphohaematopoietic engraftment. Clin Exp Immunol 154: 270-284
  • Schmidt MR, Appel MC, Giassi LJ, Greiner DL, Shultz LD, Woodland RT. 2008. Human BLyS facilitates engraftment of human PBL derived B cells in immunodeficient mice. PLoS One 3: e3192
  • Simpson-Abelson MR, Sonnenberg GF, Takita H, Yokota SJ, Conway TF, Jr., Kelleher RJ, Jr., Shultz LD, Barcos M, Bankert RB. 2008. Long-term engraftment and expansion of tumor-derived memory T cells following the implantation of non-disrupted pieces of human lung tumor into NOD-scid IL2Rgamma(null) mice. J Immunol 180: 7009-7018
  • Yamamoto T, Kaizu C, Kawasaki T, Hasegawa G, Umezu H, Ohashi R, Sakurada J, Jiang S, Shultz L, Naito M. 2008. Macrophage colony-stimulating factor is indispensable for repopulation and differentiation of Kupffer cells but not for splenic red pulp macrophages in osteopetrotic (op/op) mice after macrophage depletion. Cell Tissue Res 332: 245-256


Institutional Committees (The Jackson Laboratory)

Genetic Resources Committee
Human Subjects IRB, Chair
RAF Advisory Committee
Research Grants Committee