Charles D. Mills, Ph.D.
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PROFILE
I am best known for my discovery of M1 and M2 macrophages.
M1 and M2-type macrophages initiate and direct virtually all
other immune responses including stimulating Th1 or Th2-type
responses, respectively. This fundamental discovery has
revolutionized immunology and opened up wholly new
approaches for the immunotherapy of cancer, autoimmunity,
atherosclerosis and other diseases.
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PROFESSIONAL HIGHLIGHTS
 M1/M2 Macrophages
M1 and M2 are macrophages that influence inflammation and health in
opposing ways. M1 is a “killer” response and M2 is a “repair” response.
The M1/M2 concept also showed that Innate Immunity directs Adaptive
Immunity: A sea change in understanding how immune systems operate.
The M1/M2 discovery was included in the 2013 100 Year Timeline of
Medical Discoveries (shown). There is now explosive interest in
macrophages/innate immunity in medicine.
 Published Over 50 Articles in Immunology, Cancer, Autoimmune
Diabetes and Wound Healing
Investigations in my laboratories have resulted in influential publications
in several medical disciplines. My central interest is cancer, and my
cancer studies helped lead to the discovery of M1/M2 macrophages.
Importantly, M2-type macrophages predominate in human cancer, and
promote tumor growth. In turn, there is enormous potential in modulating
these M2 macrophages into M1 killer macrophages
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 Demonstrated That Cytolytic T Cells Can Eliminate Cancer
My postdoctoral studies at the Trudeau Institute showed that cytolytic T
cells are a key anticancer defense. These studies stimulated NIH clinical
cancer immunotherapy trials.
American Association of Immunologists
100 Year Timeline of Medical Discoveries
June 2000
M1/M2 subtypes of macrophages are identified
Mills, CD, et al. J. Immunol. 164(12): 6166-6173
Charles D. Mills and Klaus Ley
J. Innate Immunity. 2014. 6:716
Photo Credit S. Karger AG Basel
Online Timeline:
http://aai.org/timeline/digital-timeline/
 Founded BioMedical Consultants
I use my broad biomedical and creative expertise to help companies create new immunotherapies. For example, I
am on the Scientific Advisory Board of Biothera that is using a yeast-derived product to boost the immune system
against cancer in humans. (Phase II and III Clinical Cancer Trials).
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ACADEMIC AND BUSINESS EXPERTISE
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Cancer Immunology and Immunoregulation
Macrophages/Innate Immunity
Laboratory Leader/Principal Investigator
Vision: Promising Clinical Avenues for Immunotherapy
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SPECIFIC BIOMEDICAL EXPERTISE
 Immunology: Cell Culture; In Vivo and In Vitro Immunologic Analysis (e.g., Cell-Based Assays, Flow Cytometry,
ELISA, Adoptive Cell Transfer); Surgery; Histochemistry; Immunochemistry
 Cancer: Tumor Implantation and Excision; Analysis of Intratumor Environment; Cancer Inhibition Assays
 Vaccination: Adjuvant and Cell-Based Prophylaxis and Immunotherapy of Cancer
 Diabetes: In Vivo Modulation of the Immune System to Decrease Diabetes and Improve Islet Cell Transplantation
 Biochemistry: Protein Isolation and Modification; Biochemical Analysis
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PROFESSIONAL EXPERIENCE
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Founder
Designed Portable Solar Device
Associate Professor of Surgery
Assistant Professor of Surgery
Post Doc with Robert North
BioMedical Consultants
Self - Employed
University of Minnesota
Brown University
Trudeau Medical Institute
Marine, MN
Marine, MN
Minneapolis, MN
Providence, RI
Saranac Lake, NY
2007-Present
2006-2007
1991-2006
1985-1990
1980-1985
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EDUCATION
 Ph.D., Immunology /Biochemistry University of Chicago
Chicago, IL
1980
 B.S, Biology/Biochemistry
Syracuse University
Syracuse, NY
1974
 High School
Deerfield Academy
Deerfield, MA
1970
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TEACHING AND OTHER EXPERIENCE
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Taught medical immunology and mentored undergraduate and graduate students in my laboratories
Invited speaker and organizer at scientific meetings in the U.S. and abroad
NIH Immunology and Surgery Study Sections Reviewer
Editor and reviewer for medical journals (Nature, J. Exp Med., J. Immunol., Front. Immunol. J. Mol. Gen. Med.)
Host Editor for research topic at Frontiers in Immunology about M1/M2 macrophages
Medical School Admissions Committee: University of Minnesota
Animal Care Committee: Brown University and Minnesota
Head, Radiation Safety: Trudeau Medical Institute
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HOBBIES AND FAMILY
 History of science; basketball; water skiing and vegetable gardening.
 Coached men’s and women’s basketball for eight years.
 I have 2 children:
Charlie: A sophomore in college studying
automotive design and engineering.
Audrey: A chemistry major, and varsity basketball and track in college.
She is studying medicine at Johns Hopkins University.
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SELECTED PUBLICATIONS
 The following recent papers describe the central role of macrophages/innate immunity in immunity, and
how to modulate these responses against cancer and other diseases
Mills, C.D., L.L. Lenz, R. Harris. 2015. A Breakthrough: Macrophage-Directed Cancer Immunotherapy. Ca.
Research. (Invited Review, submitted August 18).
Mills, C.D. 2015. Anatomy of a Discovery: M1 and M2 Macrophages. Frontiers in Immunol. 05 May,
DOI.org/10.3389/fimmu.2015.00212.
Mills, C.D., R.A. Harris, J.D. Gregori. 2015. Cancer from your Neighbor: Adaptive Immunity to the Rescue.
Cell. 161. Link: http://www.cell.com/cell/comments/S0092-8674(15)00243-3
Mills, C.D., K. Ley, K. Buchmann, J. Canton. 2015. Sequential Immune Responses: The weapons of
immunity. J. Innate Immun. DOI:10.1159/000380910
Mills, C.D., L.L. Lenz, K. Ley. 2015. Macrophages at the fork in the road to health and disease. Frontiers in
Immunol. Doi:10.3389/fimmu.2015.00059
Mills, C.D., A.C. Thomas, L.L. Lenz, M. Munder. 2014. Macrophage: SHIP of Immunity. Frontiers in
Immunol. doi: 10.3389/fimmu.2014.00620
Mills, C.D., K. Ley. 2014. M1 and M2 Macrophages: The Chicken and the Egg of Immunity. J. Innate
Immun. 6:716.
 A recent newspaper article describes my research in laymen’s terms.
Immunologists tune in to Marine man’s discovery. 2014. Country Messenger. Osceola Sun, Publisher.
http://www.presspubs.com/messenger/news/article_887ede30-3db7-11e4-a765-eb6e0a0a57e4.html
Santoni M., S. Cascinu, C.D. Mills. 2014. Altering macrophage polarization in the tumor environment: the
role of response gene to complement 32. Cell. Molec. Immunol. Doi:10.1038/cmi.
Mills, C.D. 2012. M1 and M2 Macrophages: Oracles of Health and Disease. Critical Reviews in Immunol.
32:463.
Hering, B.J., M. Wijkstrom, M.L. Graham, M. Hardstedt, T.C. Asheim, T. Jie, J.D. Ansite, M. Nakano, J.
Cheng, W. Li, K. Moran, U. Christians, C. Finnegan, C.D. Mills, D.E. Sutherland, P. Bansal-Pakala,
M.P. Murtaugh, N. Kirchof, H.J. Schuurman. 2006. Prolonged diabetes reversal after intraportal
xenotransplantation of wild-type porcine islets in immunosuppressed nonhuman primates. Nature
(Med) 12:301.
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Mills, C.D. 2001. Macrophage arginine metabolism to ornithine/urea or nitric oxide/citrulline: A life or death
issue. Critical Reviews in Immunol. 21:399.
 The following paper describes the discovery of M1/M2 macrophages (cited in over 800 publications).
Mills, C.D., K. Kincaid, J. A. Alt, M. J. Heilman, A. H. Hill. 2000. M-1/M-2 Macrophages and the Th1/Th2
Paradigm. J. Immunol. 164:6166.
Medot-Pirenne, M., M.J. Heilman, M. Saxena, P.E. McDermott, C.D. Mills. 1999. Augmentation of an
antitumor cytotoxic T lymphocyte response in vivo by inhibition of "suppressor" macrophage nitric
oxide. J. Immunol. 163:5877.
Wahlsten, J.L., C.D. Mills, S. Ramakrishnan. 1998. Antitumor response elicited by a superantigentransmembrane sequence fusion protein anchored onto tumor cells. J. Immunol. 161:6761.
 The following paper and others below established that macrophage nitric oxide is centrally involved in
diabetes and is a major barrier to islet (cell) transplantation.
Stevens, R.B., D.E.R. Sutherland, J.D. Ansite, M. Saxena, T.J. Rossini, B.K. Levay-Young, B.J. Hering,
C.D. Mills. 1997. Insulin down regulates the iNOS pathway: Nitric Oxide as cause and effect of
diabetes? J. Immunol. 159:5329.
Shearer, J.D., J.R. Richards, C.D. Mills, M.D. Caldwell. 1997. Differential regulation of macrophage
arginine metabolism: a proposed role in wound healing. Amer. J. Physiology. 272:E181.
Stevens, R.B., J.D. Ansite, C.D. Mills, A. Lokeh, T.J. Rossini, M. Saxena, R.R. Brown, D.E.R.Sutherland.
1996. Nitric oxide mediates early dysfunction of rat and mouse islets following transplantation.
Transplantation. 61:1740
Stevens R.B., J.D. Ansite, A. Lokeh, T.J. Rossini, C.D. Mills, D.E.R. Sutherland. 1995. Expression of
intrahepatic inducible nitric oxide synthase mRNA correlates with production of nitric oxide during
intraportal isogeneic and allogeneic rat islet transplantation. Transplant. Proc. 27:615.
Ansite, J.D., R.B. Stevens, A. Lokeh, R.R. Brown, M. Saxena, C.D. Mills, D.E.R. Sutherland. 1995.
Administration of NG-monomethyl-l-arginine in a marginal renal subcapsular mouse islet transplant
model. Transplant. Proc. 27:605.
Mills, C.D., R.B. Stevens, D.E.R. Sutherland. 1994. Nitric Oxide in autoimmunity and cell transplantation.
Transplant Proc. 26:3348.
Stevens, R.B., J.D. Ansite, M. Saxena, C.D. Mills, D.E.R. Sutherland. 1994. Administration of NGmonomethyl-arginine (l-nmma) in a mouse islet transplant model. Transplant. Proc. 26:3363
Stevens, R.B., J.D. Ansite, T.J. Rossini, D.E.R. Sutherland, C.D. Mills. 1994. 15-Deoxyspergualin (DSG)
Inhibits Nitric Oxide Production In The BB/W Rat: Mechanism For Inhibition Of Islet Cell
Dysfunction In Diabetes And Transplantation. Transplant. Proc. 26:3370.
 The following paper provided the biochemical basis for why intratumor macrophages can inhibit or
promote tumor growth. These and other seminal studies in wounds (below) lead to the discovery of
M1/M2 macrophages.
Mills, C.D., J.D. Shearer, R. Evans, M.D. Caldwell. 1992. Macrophage arginine metabolism and the
inhibition or stimulation of cancer. J. Immunol. 149:2709.
Mills, C.D., J.D. Shearer, M.D. Caldwell. 1992. Macrophage arginine metabolism via nitric oxide synthase
or arginase equates with immunosurveillance or immunostimulation of cancer. In: Biology of Nitric
Oxide. S. Moncada, M.A. Marletta, and J.B. Hibbs, Eds., London; Chapel Hill: Portland Press.
Mills, C.D. 1991. Molecular basis for "suppressor" macrophages: arginine metabolism via the nitric oxide
synthetase pathway. J. Immunol. 146:2719.
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Mills, C.D., V.E. Pricolo, J.E. Albina, M.D. Caldwell. 1991. Concomitant macrophage activation and
fibroblast/lymphocyte inhibition by wound fluid: the "arginine-deficiency of inflammation" is partial
explanation. In: Clinical and Experimental Approaches to Dermal and Epidermal Repair: Normal and
Chronic Wounds. A. Barbul, Ed., New York: Wiley-Liss. Pages 193-203.
Gartner, M.H., J.D. Shearer, D.F. Bereiter, C.D. Mills, M.D. Caldwell. 1991. Wound fluid amino acid
concentrations regulate the effect of epidermal growth factor on fibroblast replication. Surgery
110:448.
Pricolo, V.E., M.D. Caldwell, B. Mastrofrancesco, C.D. Mills. 1990. Modulatory activities of wound fluid
on fibroblast proliferation and collagen synthesis. J.Surg. Res. 48:534.
Albina, J.E., C.D. Mills, M.D. Caldwell. 1990. Alterations in macrophage physiology associated with the
metabolism of L-arginine through the oxidative L-arginine deiminase pathway. In: Nitric Oxide From
L-Arginine: A Bioregulatory System. S. Moncada and E.A. Higgs, Eds., New York: Excerpta Medica.
Pages 243-248.
Albina, J.E., C.D. Mills, W.L. Henry, M.D. Caldwell. 1990. Temporal expression of different pathways of
L-arginine metabolism in healing wounds. J. Immunol. 144:3877.
Albina, J.E., M.D. Caldwell, W.L. Henry, C.D. Mills. 1989. Regulation of macrophage physiology by Larginine. Role of the oxidative L-arginine deiminase pathway. J. Immunol. 143:3641.
 The following paper at Brown University was the first describing the unique ability of macrophages to
metabolize Arginine to Nitric Oxide or Ornithine, and was the “seed” that lead to the discovery of
M1/M2 macrophages.
Albina, J.E., M.D. Caldwell, W.L. Henry, C.D. Mills. 1989. Regulation of macrophage functions by Larginine. J. Exp. Med. 169:1021.
Mills, C.D., M.D. Caldwell, D.S. Gann. 1989. Evidence of a transient "window" of immunodeficiency
following trauma of rats. J. Clin. Immunol. 9:139.
Zimbler, A., C.D. Mills, M.D. Caldwell. 1988. The effects of diabetic rat wound fluid on fibroblast growth.
Surg. Forum 39:629.
Albina, J.E., C.D. Mills, A. Barbul, C.E. Thirkell, W.L. Henry, B. Mastrofrancesco, M.D. Caldwell. 1988.
Arginine metabolism in wounds. Amer J. Physiol. 254: E459.
 The following paper and others at the Trudeau Institute established that cytolytic T cells are a key
antitumor defense, and showed that these cells are shut off by T regulatory cells during tumor growth.
Mills, C.D., R.J. North. 1983. Expression of passively transferred immunity against an established tumor
depends on generation of cytolytic T cells in the recipient. Inhibition by suppressor T cells. J. Exp.
Med. 157:1448.
Mills, C.D., R.J. North. 1985. Ly-1+2-suppressor T cells inhibit the expression of passively transferred antitumor immunity by suppressing the generation of cytolytic T cells. Transplantation 39:20
North, R.J., E.S. Dye, C.D. Mills, J.P. Chandler. 1982. Modulation of antitumor immunity-immunobiologic
approaches. Springer Sem. Immunopath. 5:193.
North, R.J., E.S. Dye, C.D. Mills. 1982. T cell-mediated negative regulation of concomitant antitumor
immunity as an obstacle to adoptive immunotherapy of established tumors. In: The Potential Role of
T Cells in Cancer Therapy. A. Fefer and A. Goldstein, Eds., New York: Raven Press. Pages 65-78.
Mills, C.D., R.J. North, E.S. Dye. 1981. Mechanisms of anti-tumor action of Corynebacterium parvum. II.
Potentiated cytolytic T cell response and its tumor-induced suppression. J. Exp. Med. 154:621.
Dye, E.S., R.J. North, C.D. Mills. 1981. Mechanisms of anti-tumor action of Corynebacterium parvum. I.
Potentiated tumor-specific immunity and its therapeutic limitations. J. Exp. Med. 154:609.