MHC Class I Loss and Cancer Immune Escape

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Federico Garrido
Departamento de Analisis Clinicos e Inmunologia,
Hospital Universitario Virgen de las Nieves,
Facultad de Medicina
Universidad de Granada
Granada, Spain

ISSN 0065-2598     ISSN 2214-8019 (electronic)

Advances in Experimental Medicine and Biology
ISBN 978-3-030-17863-5    ISBN 978-3-030-17864-2 (eBook)
https://doi.org/10.1007/978-3-030-17864-2

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Acknowledgments

The work presented in this book summarizes decades of research focused on

analyzing the expression of histocompatibility antigens in mouse and human
tumors (H-2 and HLA) in the Dept. of Analisis Clinicos e Inmunologia at
Hospital Universitario “Virgen de las Nieves” in Granada, Spain. I would like
to acknowledge all the members of the research team for their contributions.
Most importantly, I would like to thank the cancer patients in our hospital and
from different European clinical and research institutions who gave their con-
sent to analyze their tumor samples. I would like to mention Dr. Matias Perez
who started to work with me in 1979 and initiated the mouse work at the
beginning of the 1980s by inducing experimental sarcomas with methylcho-
lanthrene and typing different tumor clones for H-2 antigens. One of these
tumors, the GR9, and all the different tumor clones obtained have been stud-
ied for more than 20 years by him and other researchers in our group, Ignacio
Algarra, Jose Juan Gaforio, and Angel Garcia Lora. These studies have gener-
ated important discoveries. We have learned a lot and are still obtaining new
data from the GR9 tumor model. Dr. Teresa Cabrera set up the immunohisto-
logical laboratory and developed tumor microdissection techniques to study
HLA expression in human tumor tissues. She selected a large variety of
monoclonal antibodies that recognize different HLA antigens in tumor tis-
sues and participated in the International HLA workshop in France where the
“HLA and cancer” component was introduced for the first time. Dr. Francisco
Ruiz-Cabello applied different molecular and flow cytometry techniques to
identify and characterize molecular mechanisms responsible for HLA altera-
tions. He and his team, including Drs. Pilar Jimenez and Isabel Maleno, made
important contributions to the field. Dr. Miguel Angel Lopez-Nevot in the
early days focused on melanoma and on genomic HLA typing of tumors,
helping to develop the immunofluorescence techniques with anti-HLA mono-
clonal antibodies. Dr. Natalia Aptsiauri brought her enormous international
experience to our team, which she had gained from different USA labs, and
collaborated with me in writing different reviews and taking strategic deci-
sions in our work. All of them currently are holding important academic and
research positions at the University of Granada and Jaen, as well as in the
Tumour Tissue Biobank of the Hospital Universitario Virgen de las Nieves.
I also would like to thank clinical collaborators from our University
Hospital, including Dr. Angel Concha, the head of the Pathology Dept. who
was directly involved in providing cryopreserved tumor tissues of different
origin, for the productive cooperation that we have had for many years in

v
vi Acknowledgments

d­ ifficult times; Dr. Miguel Tallada and Dr. Jose Manuel Cozar (Urology
Dept.), Drs. Antonio Cueto and Abel Sanchez-Palencia (Thoracic Surgery),
Dr. Javier Gutierrez and Dr. Antonio Ferron (General Surgery), Prof.
Francisco Esteban and Dr. Jose Salinero (Otorhinolaryngology Dept.), Prof.
Alfonso Herruzo (Gynecology Dept.), and Prof. Salvio Serrano (Dermatology),
for cooperating with us at different time periods providing fresh tumor tissue
samples of different origin; and the pathologist Dr. Miguel Angel Piris from
Madrid and the immunologists Dr. Francisco Real and Prof. Miguel Lopez-­
Botet from Barcelona for helping to design new approaches in the HLA and
cancer field.
I must also mention important and diverse international cooperations that
we developed over the years starting with the collaboration with Dr. Peter
Stern from the Patterson Institute in Manchester, England. We published two
highly quoted reviews in 1993 and 1997, which summarize the work carried
out in Granada and Manchester in the 1980s and 1990s in relation to HLA
expression in human tumors. Dr. Eva Klein from the Karolinska Institute has
been interested in this area of research from the very beginning and orga-
nized, together with Dr. Hilliard Festenstein from the London Hospital, the
first meeting on “Histocompatibility Antigens in Tumors” that took place in
Granada in 1985 (see J of Immunogenetics Vol.13, n°2/3, 1986). Drs. Klass
Karre and Hans-Gustaf Ljunggren from the same institution collaborated
with us in defining the complexity of the different clones of the mouse GR9
fibrosarcoma. Dr. Soldano Ferrone, a pioneer in the HLA and cancer field,
collaborated with us when he was working in New York State, in Pittsburg,
and later in Harvard University in Boston. Dr. Francesco Marincola at the
NIH (USA) helped us to analyze in detail progressing and regressing meta-
static lesions in mixed responder melanoma patients. Together with Dr.
Marcel Tilanus and Manita Feenstra from Utrecht, we developed novel tech-
niques to define HLA haplotype losses in human tumors and participated in
the HLA workshops. With another leading scientist in our field, Dr. Barbara
Seliger from Halle in Germany, we collaborated in the investigations of “the
antigen presentation machinery” in tumor cells. I have to mention an impor-
tant collaboration with Drs. Dirk Schadendorf and Annette Paschen
(Mannhein and Essen) in defining new molecular mechanisms responsible
for HLA alterations in melanoma. With Dr. Gustav Gaudernak and his team
at the Norwegian Radium Hospital in Oslo, we have a long-term collabora-
tion studying HLA class I loss in tumor immune escape and cancer progres-
sion. Dr. Thierry Boon and Pierre Coulie from the Ludwig Institute in
Brussels, who pioneered the identification of tumor antigens recognized by T
lymphocytes, cooperated with us in defining HLA-I-associated tumor
immune escape mechanisms in patients receiving peptide immunotherapy.
With Drs. Alex Knuth (Mainz), Catia Traversari (Milan), and Jesper Zeuthen
(Copenhagen), we worked closely defining new molecular mechanisms of
HLA alteration in melanoma. With Drs. Graham Pawelec (Tubingen) and
Steve Marsh (London), we collaborated in an EU project in which a European
melanoma cell bank (ESTADB) has been developed.
I want to highlight the period I spent as a postdoctoral fellow in the London
Hospital Medical College under the supervision of Dr. Hilliard Festenstein.
Acknowledgments vii

He helped me to start a new project in his lab typing H-2 antigens in mouse
tumors in 1974–1975. I learned immunogenetics of the H-2 system and
applied this knowledge to the altered HLA expression observed in tumor
cells. This was the beginning of my journey in the field of “MHC and cancer.”
Drs. Dominique Charron (Paris) and Renee Fauchet (Nantes) introduced for
the first time the “HLA and cancer” component in the XII International
Histocompatibility workshop in 1996. I would also like to thank the Spanish
Medical Research Council (Fondo de Investigaciones Sanitarias, FIS) for
financing our research since 1981 and “Ramon Areces Foundation” in Spain
for financing the “HLA and Cancer” meetings organized in Granada in 1998
and 2011. I would like to thank many different PhD and postdoctoral students
that worked in our group at Hospital Universitario Virgen de las Nieves in
Granada since 1981: Javier Martin, Concha Delgado, Rosario Oliva, Susana
Pedrinaci, Maria Luisa Garrido, Antonio Garrido, Abelardo Caballero,
Maximino Redondo, Julia Canton, Alfonso Serrano, Rosa Mendez, Jose
Maria Romero, Monica Bernal, Teresa Rodriguez, Francisco Perea, Luis
Miguel Real, Rafael and Javier Carretero, Irene Romero, Cristina Garrido,
Ana Belen del Campo, Isabel Maleno, and Greta Garrido from the Molecular
Immunology Institute of Havana, Cuba. All made important discoveries pub-
lished in leading international journals. I would specifically thank Dr. Monica
Bernal for her help in making different figures of this book, Dr. Fran Perea for
the images of tissue immunohistochemistry, Dr. Teresa Rodriguez and Maria
Jose Olivares for helping me with the reference list, and Drs. Natalia Aptsiauri,
Francisco Ruiz-Cabello, and Angel Garcia Lora for the critical review of the
book. Finally, I would like to thank my family, my wife, Antonia Collado,
who has been always by my side in difficult moments and helped me with my
research for many years working in the Research Unit of our Hospital, and
my three daughters, Pilar, Carolina, and Nona, who missed me many times
when I was travelling to different places and meetings.
This book summarizes the progress made in the field of “MHC and Cancer
Immune Escape” and the contributions made by many scientists all over the
world. Nevertheless, there is still a long way before we fully understand the
“stealth technology” used by cancer cells to escape and find new methods to
fight it that can be applied in the clinic. MHC was discovered in experiments
with tumor transplantation in mice. The impact of HLA tissue typing in
human transplantation helped to improve our knowledge about this complex
genetic system. Now MHC/HLA returns to cancer owing its important role in
tumor rejection and escape. The MHC system did not evolve to fight cancer
but is no doubt playing a pivotal role. Perhaps, MHC is now “repaying” to
cancer cells for being discovered. It is likely that “MHC and cancer” will be
travelling together for the years to come.
Contents

1 Introduction��������������������������������������������������������������������������������������   1
1.1 The Early Days of Tumour Immunology����������������������������������   3
1.2 The Major Histocompatibility Complex (MHC):
The HLA and the H-2 System��������������������������������������������������   4
1.3 Antigen Processing and Presentation via HLA
Class I/II Molecules������������������������������������������������������������������   8
1.4 The Expression of HLA-I and II Molecules
in Normal Tissues ��������������������������������������������������������������������   8
References������������������������������������������������������������������������������������������ 10
2 MHC/HLA Class I Loss in Cancer Cells �������������������������������������� 15
2.1 MHC/HLA Class I Loss in Primary Tumors���������������������������� 16
2.1.1 H-2/HLA-I Heterogeneity�������������������������������������������� 17
2.1.2 Altered HLA-I Tumor Phenotypes�������������������������������� 19
2.1.3 The Transition from HLA-I Positive to HLA-I
Negative Tumors Induced by T Cell Immune
Selection: Impact on Tumor Tissue Architecture���������� 27
2.1.4 HLA Class I Loss in Different Tumor Tissues�������������� 31
2.1.5 Non Classical HLA Class I Molecules in Tumors�������� 54
2.2 MHC/HLA Class I Expression in Metastasis �������������������������� 56
2.2.1 H-2 Class I Expression in Spontaneous
Metastases in Mice�������������������������������������������������������� 57
2.2.2 HLA Class I Expression in Human Metastasis������������ 59
2.2.3 MHC/HLA Class I Expression and Metastatic
Dormancy���������������������������������������������������������������������� 62
References������������������������������������������������������������������������������������������ 63
3 HLA Class-I Expression and Cancer Immunotherapy���������������� 79
3.1 Reversible “Soft” Versus Irreversible “Hard”
Molecular HLA-I Lesions: Implications
for Cancer Immunotherapy ������������������������������������������������������ 80
3.2 Recovery of HLA-I Antigen Expression in Tumors
with “Hard Lesions”: A Challenge for the Future�������������������� 84
References������������������������������������������������������������������������������������������ 87

ix
x Contents

4 HLA Class-II Expression in Human Tumors�������������������������������� 91

References������������������������������������������������������������������������������������������ 94

Looking into the Future���������������������������������������������������������������������������� 97

Concluding Remarks �������������������������������������������������������������������������������� 99

Index���������������������������������������������������������������������������������������������������������� 101
 Introduction
1

Abstract Peter Gorer discovered the Histocompatibility sys-

This chapter focuses on the discovery of the tem (H-2) in mice in 1935 in experiments with
Major Histocompatibility Complex (MHC) in allogeneic tumor transplantation. The key obser-
mice (H-2) and in humans (HLA), and on the vation was that tumors fail to growth when trans-
role played by the International HLA Workshops planted into genetically distinct host of the same
in the analysis and characterization of this com- species. The antibodies generated in these alloim-
plex genetic system. The early days of Tumour munizations were used for MHC typing of red
Immunology and the importance of the defini- blood cells in inbred strains of mice using a
tion of Tumour Associated Transplantation complement-­ dependent cytotoxicity test (Gorer
Antigens (TATA) are also discussed. Today we 1936, 1937; Gorer and Mikulska 1954; Gorer and
know that tumour cells can be killed by T lym- O’Gorman 1956). This test was used years later to
phocytes by recognizing tumour antigenic pep- discover and define the HLA system in humans by
tides presented by MHC molecules and they can testing leukocytes with alloimmune sera obtained
also escape this recognition by losing the from individuals after blood transfusion (Dausset
expression of MHC molecules. This important 1954) and from pregnant women (van Rood 1962).
phenomenon has been profoundly studied for I recommend reading the article “Seeds of time:
many years both in my lab in Granada and in Fifty years ago Peter A. Gorer discovered the H-2
other laboratories. The results of this research complex” written by Jean Klein (1986), the publi-
have important implications for the new genera- cation by Bernard Amos on “Recollections of Dr
tion of cancer immunotherapy that boosts T cell Peter Gorer” (1986) and the book “H-2 antigens:
responses. A historical perspective of major dis- genes, molecules, function” edited by Shella David
coveries is presented in this chapter, with the (1987) after the meeting he organized in the
names of the scientists that have made a signifi- Jackson Lab in Bar Harbor (Maine, USA) to com-
cant contribution to the enormous progress memorate the 50th anniversary of the discovery of
made in the field of Tumour Immunology. the H-2 system (see also Snell 1986).
In 1975–1976 mouse tumors were typed for
Keywords H-2 antigens in the laboratory of Hilliard
MHC · HLA · H-2 · Major histocompatibility Festenstein at the London Hospital Medical
complex · HLA heavy chain · Beta2 micro- College using mouse alloantisera, defining pri-
globulin · Tumor immunology · Tumor vate and public H-2 specificities (Garrido et al.
escape · HLA workshops · Antigen 1976, 1977, 1979; Schmidt et al. 1979). The
processing key finding was that the MHC class I profiles

© Springer Nature Switzerland AG 2019 1

F. Garrido, MHC Class-I Loss and Cancer Immune Escape, Advances in Experimental Medicine
and Biology 1151, https://doi.org/10.1007/978-3-030-17864-2_1
2 1 Introduction

obtained when typing mouse tumors were not Smith et al. 1989; Bodmer et al. 1993; Garrido
the same as those corresponding to autologous/ et al. 1995; Ruiz-Cabello and Garrido 2002).
syngeneic lymphoid cells. Extra reactivities as Viruses also use similar escape strategies (Schrier
well as the absense of expected reactions (H-2 et al. 1983; Alcami and Koszinowski 2000).
class I losses) were discovered. The former Although this mechanism was described in
were, most likely, due to the anti-viral antibod- mouse and human tumors many years ago
ies present in the mouse alloantisera. The latter (Garrido et al. 1979, 1997; Festenstein 1987;
were unexpected H-2 reactivity patterns that Garrido and Klein 1991; Marincola et al. 2000),
could not be clearly explained at the time. This it has been ignored for decades. However, a
work was the beginning of a history with revival of the topic “HLA and cancer immune
important implications in Tumour Immunology escape” is now in progress with new reports by
and Clinical Oncology (Festenstein and different groups involving HLA in tumor escape
Schmidt 1981; Festenstein 1987). (Lampen and van Hall 2011; Chowell et al. 2017;
HLA tumor typing in human melanoma and McGranahan et al. 2017; Sucker et al. 2017;
colorectal carcinoma was performed in the 80s in Grasso et al. 2018; Aptsiauri et al. 2018). The old
tumor tissues using anti-HLA monoclonal anti- idea that the immune system is capable to con-
bodies that recognize monomorphic determinants stantly survey and destroy transformed cells in
(Ruiter et al. 1982; Natali et al. 1983; Csiba et al. our body (Burnet 1957, 1970, 1971; Thomas
1984). The HLA class I positive staining of the 1959) was confirmed when a particular tumor
tumor stroma was always used as an internal con- escape mechanism had been defined in a given
trol (Ruiter et al. 1984, 1986; Natali et al. 1984; tumor (Villunger and Strasser 1999).
Lopez-Nevot et al. 1986; Moore et al. 1986; This book analyzes the relevance of MHC/
Cabrera et al. 2003). It was evident from the very HLA expression for the immune escape in primary
beginning that HLA typing of tumor tissues did tumors and metastasis, including experimental
not match that of autologous peripheral blood data obtained in chemically induced sarcomas, as
lymphocytes of a particular cancer patient. The well as in human tumors. These data are the result
absence of HLA class I expression was detected of many years of work in the Clinical Laboratory
with anti-HLA MoAbs directed against common of the “Hospital Universitario Virgen de las
HLA A,B,C determinants, against the β2 micro- Nieves” in Granada, Spain. During this long period
globulin or against particular HLA class I alleles of time, which started in the late 70s, five scientific
(Lopez-Nevot et al. 1989; Natali et al. 1989). meetings took place in Granada (Spain) on the
After decades of research doing HLA typing of topic “HLA and Cancer”. The first one was held in
tumor tissues by different groups including our- 1985 with the title: “Histocompatibility Antigens
selves, it has become accepted that altered tumor on Tumours”. A special issue of the Journal of
HLA class I expression is a major finding due to Immunogenetics published a set of papers on this
its high frequency and its relevance in cancer topic presented during the meeting (see Butcher G,
immune escape (Garrido et al. 1993; Marincola Journal of Immunogenetics 1986 Vol 13, N° 2/3,
et al. 2000; Garrido and Algarra 2001). ­April/June). The second one took place in 1991
There is strong evidence suggesting that HLA and was titled: “MHC molecules in normal and
class I cell surface expression in tumor cells plays neoplastic cells”. A special issue of the
a pivotal role in the recognition and destruction International Journal of Cancer reported the differ-
of nascent tumors due to its role in antigen pre- ent presentations that took place at the meeting
sentation to T lymphocytes. Loss or downregula- (see Garrido F, Int J Cancer 1991, supplement 6).
tion of MHC/HLA class I molecules is a key The third one in 1994 in the city of Jaen, near
mechanism used by tumor cells to escape from T Granada with the title of “Oncogenes, tumour
cell responses, as it has been demonstrated by antigens and MHC Molecules” and reported in
different research groups (Schmidt and volume 47 of Tissue Antigens 1996 (see Garrido
Festenstein 1982; Festenstein and Garrido 1986; 1996). The fourth one took place in 1998 in
1.1 The Early Days of Tumour Immunology 3

Granada with the title of: “Tumour Antigens and The discovery of NK cells by Rolf Kiessling
MHC molecules: Mechanism of tumour escape and colleagues at the Tumor Biology Department
from the immune system, implications for immuno- of the Karolinska Institute (Kiessling et al. 1975;
therapy” and the last one in the village of Dílar Kiessling and Wigzell 1979) opened a new field
near by Granada in October, 2011 with the title of also associated with MHC-I/HLA-I cell surface
“Cancer Immune escape: implications for immu- expression in tumors and virus infected cells.
notherapy” (Aptsiauri et al. 2012). Also the Several years later in the same Department it was
PIVAC 6 meeting (Progress in Vaccination Against reported that NK cells are activated by the
Cancer) was organized in our hospital in Granada “absence” of MHC class I molecules in the tumor
in 2006 and was dedicated to HLA and Cancer target, raising the provocative idea of the “Missing
(Aptsiauri et al. 2007a). The Granada meetings in Self hypothesis” with important implications in
Tumour Immunology had a common goal, namely, Tumor Immunology (Kärre et al. 1986; Ljunggren
to analyze the role of MHC antigens in cancer and Kärre 1986, 1990; Kärre 1997). The discov-
immune escape. ery of tumor antigens recognized by T lympho-
cytes was also an important accomplishment
made at the Ludwig Institute in Brussels by
1.1  he Early Days of Tumour
T Thierry Boon and colleagues (Boon 1983; Van
Immunology der Bruggen et al. 1991; Boon et al. 1994, 1996,
2006). They showed that “shared or unique-­
The ability of the immune system to reject mutated tumor peptides” are recognized by T
tumors in syngeneic animals was first demon- lymphocytes and restricted by the expression of a
strated in inbred mice with chemically induced particular MHC/HLA-I allele. The expression of
sarcomas (Gross 1943; Foley 1953; Baldwin the MAGE genes in melanoma and other tumors
1955; Prehn and Main 1957; Klein et al. 1960). were extensively tested for T cell responses rec-
Mice immunized with a particular sarcoma were ognizing the corresponding tumor peptides pre-
protected against the same tumor but not against sented by different HLA class I alleles (Chomez
other types. These experiments, done in different et al. 2001). All these findings clearly indicated
labs during the last century, originated the con- that MHC/HLA molecules play a crucial role in
cept of “Tumour Associated Transplantation immune responses against viral infections and
Antigens” (TATA) and also introduced the idea cancer by interacting with T and NK cell
of “Individual-Unique Tumour Antigens” receptors.
(Basombrio 1970). These discoveries were in Historically, the main objective of cancer
line with the idea proposed at the time by immunotherapy has been boosting T cell
McFarlane Burnet that the immune system can responses against tumors. This includes BCG
survey the cells of our body. He wrote: “It is an therapy in superficial bladder cancer (Morales
evolutionary necessity that there should be some et al. 1976), Interleukin-2 (Rosenberg et al.
mechanism for eliminating or inactivating such 1987), immunization with tumor peptides as
potentially dangerous mutant cells and it is pos- tumor antigens (Marchand et al. 1999), dendritic
tulated that this mechanism is of immunological cells loaded with tumor peptides (Nestle et al.
character” (Burnet 1957, 1970, 1971). A major 1998), the use of biological response modifiers
breakthrough in immunology was the discovery such as Polysacharide K (PSK) (Nakazato et al.
that T lymphocyte-mediated cytotoxicity (CTL) 1994; Nio et al. 1991) and adoptive cell transfer
is restricted by MHC molecules (Zinkernagel (ACT) using in vitro expanded tumor infiltrating
and Doherty 1974, 1979, 1997; Zinkernagel lymphocytes (TIL) in metastatic melanoma
1996) and that virus antigens are recognized as (Rosenberg et al. 2011; Andersen et al. 2016).
small peptides inside the structure of the MHC Today cancer immunotherapy is reaching a peak
molecules (Bjorkman et al. 1987; Townsend with novel therapies using antibodies that target
et al. 1986). molecules involved in the regulation of T cell
4 1 Introduction

cytotoxicity (Allison et al. 1982, 1995; Leach International HLA Workshop was organized by
et al. 1996; Kvistborg et al. 2014; Buferne et al. Bernard Amos at Duke University in USA in 1964
2015) reinforcing the idea that T lymphocytes are to exchange anti-HLA sera and to analyze the dif-
playing a major role in recognizing and destroy- ferent reactivities obtained from testing the allo-
ing cancer cells (Svane et al. 1999; Boesen et al. immune sera against leucocytes using a
2000; Romero and Coulie 2014). However, only complement-mediated cytotoxicity test. It was
a proportion of cancer patients benefit from this followed by a series of workshops that were orga-
therapy and demonstrate tumor regression, while nized by pioneers in the HLA field: in 1965 John
others develop therapy-resistant progressing met- Van Rood in Leiden (Holland), in 1967 Rugero
astatic tumor lesions. The reason and underlying Cepellini in Torino (Italy), in 1970 Paul Terasaky
mechanisms of these clinical variations are not in Los Angeles (USA), in 1972 Jean Dausset in
known. Hence, it is very important to define in Paris (France), in 1973 Kisseyer Nielsen in Aarhus
the future studies the different escape strategies (Denmark), in 1977 Walter Bodmer in Oxford
used in primary and metastatic tumor lesions and (England) and many more, as an example of an
personalized suitable therapy in each patient. It is International cooperation between scientists to
becoming evident that tumors can be rejected, define and understand the structure and function
escape or be kept in the state of immune medi- of such a complex biological system. A single
ated “equilibrium or dormancy”, as it was pro- laboratory was not able to analyze the multiple
posed by Bob Schreiber in the “immunoediting” different reactivities obtained with many different
concept (Dunn et al. 2002, 2004). These different sera that were tested againts a large variety of leu-
tumor/host interactions are very much related to cocytes as targets. It was necessary to exchange
the level of MHC/HLA antigens expressed by the these reagents and to compare the patterns of
tumor cells (Aptsiauri et al. 2007b; Garrido et al. reactivities by different HLA labs. These work-
2016). Modern cancer immunotherapy will need shops still take place and the last one was in
to monitor the HLA expression levels in tumor Standford University (CA, USA) in October
tissues in order to avoid ineffective therapies 2017, and was organized by Marcelo Fernandez-
mediated by T cells when the target peptide anti- Viña and Peter Parham. In order to understand the
gen is not present (Aptsiauri et al. 2008; Thor history of HLA and the pioneers that organized
Straten and Garrido 2016). such HLA workshops, I recommend reading the
Introduction of the 13th International HLA work-
shop report written by John Hansen (2006) that
1.2 The Major took place in Victoria (British Columbia) and in
Histocompatibility Complex Seattle (USA) in 2002. Interestingly, the organi-
(MHC): The HLA and the H-2 zation of the HLA ­workshops was used years later
System to prepare the Leucocyte Differentiation Antigens
workshops that produced the definition of the dif-
The MHC molecules in humans are called Human ferent molecules called “Clusters of
Leucocyte Antigens (HLA). The HLA molecules Differentiation” or “CDs” also with an enormous
were discovered by alloimmune interactions but impact in Haematology, Pathology and Clinical
they play a major role in antigen presentation to T Medicine. The first CD Workshop took place in
lymphocytes and in cancer immune escape. It is Paris in 1982 and was organized by Alain Bernard;
therefore necessary to briefly summarize our cur- the second one in Boston was organized by Stuart
rent knowledge about the structure and function Schlossman, and the third one in Oxford by
of HLA genes and molecules that are expressed in Andrew McMichael in September of 1986. I rec-
most nucleated cell. Jean Dausset in Paris and JJ ommend reading the introductory remarks of the
Van Rood in Leiden discovered the HLA system third CD workshop writen by Cesar Milstein
(Dausset 1958; van Rood 1962). The 1st (1987).
1.2 The Major Histocompatibility Complex (MHC): The HLA and the H-2 System 5

Today we know that the HLA system is a com- population is extremely high with 13,262 for
plex genetic structure located in the short arm of HLA class I and 4,846 for HLA class II as defined
chromosome six composed of different loci clas- today. See IPD-IMGT/HLA Database web site
sified in Class I, class II and class III. See review for updated details https://www.ebi.ac.uk/ipd/
written by Jan Klein in 2000 where this classifi- imgt/hla/stats.html.
cation was proposed for the first time (Klein and HLA class I molecules are cell surface glyco-
Sato 2000a, b). HLA genes are inherited in a proteins composed of highly polymorphic heavy
Mendelian form giving rise to the most polymor- chains encoded by genes located in the short arm
phic antigenic system that exists in humans (see of chromosome 6 covalently linked to a constant
Fig. 1.1). Within the class I region there are three small molecule i.e. the β2 microglobulin encoded
classical genes (HLA-A, B and C) and three non-­ in chromosome 15 (Fig. 1.1). The extracellular
classical genes (HLA-E, F, G). Each individual part of the heavy chain is devided into three
has a combination of six HLA-I classical HLA-I domains: α1, α2 and α3. The peptide binding
genes (loci A, B and C) in the majority of their groove is formed by folding the α1 and α2
cells. The number of HLA-I alleles in the human domains to create a cleft where the aminoacid

Fig. 1.1 Genetic structure of the Major stant light chain (β2 microglobulin) encoded in
Histocompatibility Complex in humans (The HLA sys- chromosome 15. Antigen presenting cells (APCs) express
tem) and cell surface expression of HLA I and II mol- additional 6 HLA class II molecules: DR, DP and DQ
ecules. It is located in the 6p21 region of chromosome 6. composed of two chains (α and β chains) encoded by the
Any somatic cell expresses six HLA class I alleles com- genes within the HLA region
posed of a polymorphic heavy chain (A,B,C) and a con-
6 1 Introduction

Table 1.1 Tissue distribution of HLA class I and II antigens in normal tissues
Tissue Immunolabeling Tissue Inmunolabeling
HLA-I HLA-II HLA-I HLA-II
Cells of the endocrine Urogenital System:
system:
Thyroid (follicular, para + − Kidney glomeruli ++ ++
follicular)
Pancreatic isles of Langerhans + − Kidney tubules ++ ++/−/+/−
Grastrointestinal tract: Epithelium
 Esophagus ++ −  Ureter ++ −
 Stomach + −  Bladder ++ −
 Duodenum ++ ++  Prostate ++ −
 Colon ++ −  Urethra ++ ++
 Rectum ++ − Testis
 Gall bladder + ND  Germ cell line, + −
spermatozoa
Liver Miscellaneous:
 Hepatocytes +/− − Breast epithelium
 Biliary apithelium ++ −/+/−  Ductal ++ ++
Respiratory and  Glandular ++ ++
cardiovascular systems:
Tonsillary epithelium ++ ++ Pancreas
Lung (bronchial and alveolar ++ ND Exocrine portion − −
epithelium)
Heart Ductal epitelium ++ −
 Myocardium + − Muscle
Endothelium  Skeletal +/− −
 Capillaries ++ ++  Smooth + −
 Large vessels ++ +/−/+/− Langerhans cells, ++ ++
interstitial dentritic cells
Nervous system Lymphatics ++ ++
Peripheral ++ − Fibroblasts ++ −
Central Placenta-­villous trophoblas − −
 Neurones − − Epidermis + −
(−) negative; (−/+)…… (+) weakly positive; (++) strongly positive

residues cluster in the hypervariable region (Stern For instance: normal lymphoid cells are express-
and Wiley 1994). The groove formed by these ing high amount of HLA-I, hepatocytes have
domains anchor the peptide recognized by the T lower expression and colorectal normal tissues
cell receptor (TCR). Similarly, HLA class II mol- have an intermediate level. See Table 1.1 sum-
ecules are composed of two polymorphic chains marizing the level of HLA expression in different
coded by genes located in the centromeric part of human tissues. A careful analysis of HLA-I
the HLA complex and composed of three differ- expression in different normal tissues is manda-
ent loci: HLA DR, DP and DQ (see Fig. 1.1). tory since the data that exist were published long
Most nucleated cell in the body express six time ago using only immunohistological tech-
different HLA class I molecules: two HLA-A, niques (Daar et al. 1984a). HLA-II molecules
two HLA-B and two HLA-C, but the amount of have a restricted tissue distribution since are
these molecules expressed at the cell surface vary expressed in antigen presenting cells (APCs)
significantly depending on the level of gene tran- such as dendritic cells, macrophages, B cells,
scription, transduction and epigenetic regulation. activated T lymphocytes and some epithelia, such
1.2 The Major Histocompatibility Complex (MHC): The HLA and the H-2 System 7

as cervical and colorectal that are under exoge- This interaction is regulated by a set of NK recep-
nous exposure to pathogens (Daar et al. 1984b; tors that recognize HLA-I molecules: a soub-
Cabrera et al. 1995). group of HLA-B locus products (HLA Bw4
HLA class I and II molecules have important positive alleles) and HLA-Cw molecules as two
immunological functions: the presentation of different entities, i.e. HLA-C1 and HLA-C2
foreing or altered antigens to T lymphocytes as (Moretta et al. 1996; Lanier and Philips 1996;
small peptides inside the HLA structure. Each Lanier 2005). The non-classical HLA-E is also
HLA allele binds and presents to T-cells a par- interacting with a specific NK cell receptor
ticular peptide aminoacid sequence derived from named CD94/NKG2D (Lopez-Botet and Bellon
a particular protein in a highly specific manner in 1999). NK receptors do not recognize HLA-I
the polymorphic “groove” of the molecular com- allelic differences like T cell receptor do.
plex (Rotzschke et al. 1990; Falk et al. 1991; The MHC genes and molecules in mice are
Rammensee et al. 1995, 1999). Each individual known as H-2 and are located in chromosome
carries a set of six different HLA I and six differ- seventeen. The H-2 system was discovered by
ent HLA-II alleles with a defined capacity to Peter Gorer in 1935–1936 doing tumour trans-
present antigens (peptides) to T lymphocytes. plantation in different imbread strains of mice
HLA class I interacts with the T cell receptor of (Gorer 1936, 1937). Inbred mice are homozy-
CD8 positive lymphocytes and HLA class II with gous and both chromosomes carry the same
the same structure in CD4 positive lymphocytes. genes. The commontly used laboratory mice
In addition, HLA-I molecules also have the abil- present the H-2 haplotypes H-2d (BALB/c), H-2k
ity to interact with Natural Killer (NK) cells, but (CBA/H), H-2b (C57/BL6), H-2a (A/Jax), among
in this case inhibiting their function (Fig. 1.2). others. There are two loci for class I: K and D that

Fig. 1.2 Interaction of HLA class I molecules with T HLA-I molecules expressed on the tumor cell surface
and NK cell receptors. Tumor cells and virus infected interact with NK cell receptors inhibiting their function.
cells present tumor antigens to CD8 positive T lympho- So, HLA class I molecules can activate T cell function and
cytes as nine-aminoacide tumor peptides situated inside at the same time inhibit NK cell cytotoxicity
the HLA class-I molecule structure. At the same time,