IMMUNGLOBULINS STRUCTURE and FUNCTION Immunoglobulin structure Valency, affinity, avidity Direct and indirect functions Immunoglobulin types Fc receptors Immunoglobulin transport Monoclonal and polyclonal antibodies Passive immunization Therapeutic antibodies Chimera and humanized monoclonal Ab Tumortherapy, Immunsuppression Case study (mieloma multiplex) B cell malignancies More and more therapeutic monoclonal antibodies CARs and TRUCKs

IMMUNOGLOBULINS Definition: Glycoprotein molecules that are present on B cells (BCR) or produced by plasma cells (antibodies) in response to an immunogen immunoglobulin ~ antibody

STRUCTURE heavy and light chains disulfide bonds inter-chain intra-chain disulfide bond carbohydrate CL VL CH2 CH3 CH1 VH hinge region

STRUCTURE variable and constant regions hinge region domains immunoglobulin domen variable and constant regions hinge region hinge region carbohydrate disulfide bond CH1 CL VH CH2 CH3 domains VL & CL VH & CH1 - CH3 (or CH4) oligosaccharides VL

Ribbon structure of IgG

mIg = BCR Ne legyen megtévesztő, hogy az eddigi ábrákon a könnyűlánc volt belül, itt meg kívül vannak. Ábrázolás szempontjából tök mindegy. míg – membrán horgonyzott immunoglobulin = BCR (B cell receptor), a B sejt antigénreceptora A bekarikázott rész lesz ábrázolva a következő kép alján

Hypervariable region - Complementary Determining Region (CDR) 3 CDR regions in a V domain VH & VL domains 3+3 CDR (the circled part from the previous slide)

IMMUNOGLOBULIN FRAGMENTS STRUCTURE/FUNCTION RELATIONSHIPS antigen binding Az ellenanyag molekula különböző részeihez, különböző funkciók társíthatók complement binding site placental transfer binding to Fc receptors

IMMUNOGLOBULIN FRAGMENTS STRUCTURE/FUNCTION RELATIONSHIPS papain Fab antigen binding (valence=1, monovalent) specificity determined by VH and VL domains some effector function is impaired VH VL Fab – Fragment antigen binding Fc – Fragment crystallizable  mivel kicsi a variabilitás a fargmentumok között, könnyebben kristályosítható Figyeljük meg a hasítás helyét a nehézláncokat egymással összekötő diszulfid hidakhoz képest! Fc Fc (effector functions) Fab

IMMUNOGLOBULIN FRAGMENTS STRUCTURE/FUNCTION RELATIONSHIPS pepsin F(ab’)2 bivalent antigen binding (valency=2) Fc peptides + bivalens – képes lehet precipitációra, agglutinációra (A PEPSI-t nem kell komolyan venni) Figyeljük meg a hasítás helyét a nehézláncokat egymással összekötő diszulfid hidakhoz képest! F(ab’)2 Some effektor functin is impaired. It can mediate agglutination/precipitation (other seminar)

MODEL OF ANTIGEN BINDING Egyszerű modell, hogyan lehet elképzelni az antigén kötőhelyet (kötőfelszínt) illetve az idiotípus régiót (az epitoppal reagáló rész)

EFFECTOR FUNCTION: NEUTRALIZATION neutralized pathogen / toxin Ez az effektor funkció szinte csak a variabilis domén (az antigénkötő rész) tulajdonságaitól függ. Az ellenanyag más részei (pl. az izotípus) nem befolyásolják (vagy csak közvetve – lásd a valenciánál) Immunizálás során a szervezetben képződő/bejuttatott ellenanyagok így védenek vírusfertőzésk ellen Toxinok hatását blokkolhatják Baktérium sejtek adhézióját gátolhatják az epitélsejteken toxin effect or pathogen adhesion is blocked by steric hindrance

Why do antibodies need an Fc region? detect antigen precipitate antigen block the active sites of toxins or pathogen-associated molecules block interactions between host and pathogen-associated molecules the (Fab)2 fragment can NEUTRALIZATION inflammatory and effector functions associated with cells inflammatory and effector functions associated with complement system the trafficking of antigens into the antigen processing pathways (opsonized phagocytosis) but can not activate (role of Fc region) Neutralization is an effector function related to the Fab region

EFFECTOR FUNCTIONS OF ANTIBODIES Neutralization (variable domain associated) Marking the antigens for effector functions (variáble domain and Fc dependent) phagocytosis (oppsonization) ADCC and IgE mediated mast cell activation activation of the complement system (next seminar) Fc part is recognized by Fc receptors Different antibodies have different Fc part

VARIABILITY IN DIFFERENT REGIONS OF THE IG DETERMINES IG CLASSES OR SPECIFICITY isotype allotype idiotype (Classes/subclasses) Sequence variability of H/L-chain constant regions Allelic variants Sequence variability of H and L-chain variable regions (individual, clone- specific)

HUMAN IMMUNOGLOBULIN CLASSES encoded by different structural gene segments (isotypes) IgG - gamma (γ) heavy chains IgM - mu (μ) heavy chains IgA - alpha (α) heavy chains IgD - delta (δ) heavy chains IgE - epsilon (ε) heavy chains light chain types kappa (κ) lambda (λ)

Az Fc receptors FcγR : γ  IgG binding FcεR : ε  IgE binding FcαR : α  IgA binding

Phagocytosis is ineffective/slow without opsonization Ógörög opszon: Főzött, megfelelően ízesre elkészített étel. Az étkezést kiegészítő, ízesebbé tevő étel, gyakran halétel. Mellékétel (side dish), de nem lekicsinylő értelemben. Az étkezés legkívánatosabb, legízesebb részét, vagy fűszert, sót is érthettek rajta. http://en.wikipedia.org/wiki/Opson http://www.perseus.tufts.edu/hopper/text?doc=Perseus%3Atext%3A1999.04.0063%3Aalphabetic+letter%3DO%3Aentry+group%3D1%3Aentry%3Dopson-cn Phagocytosis is ineffective/slow without opsonization

complex antigen antibodies complexed to complex antigen (A) High-affinity FcRs on the surface of the cell bind monomeric Ig before it binds to antigen. (B) Low-affinity FcRs bind multiple Igs that have already bound to a multivalent antigen.

ANTIBODY DEPENDENT CELLULAR CYTOTOXICITY (ADCC) low affinity FcR mediated NK cell / Basophyl granulocyte degranulation Bazofil granulociták az IgE/FreR mellett FcgReceptorral is rendelkeznek - hasonló folyamattal aktiválódhatnak és üríthetik granulumaikat.

MAST CELL DEGRANULATION Mast cell pre-armed with antigen specific IgE Cross-linking of IgE via bound pathogen/allergen degranulation Az IgE szabad állapotban is képes kötődni a nagy affinitású FcεR receptorhoz. A sejt ilyen állapotban várja a találkozást az antigénnel (kissé a B sejtek antigénfelismerésére emlékeztető módon FcεR kötött ellenanyag  mIg (BCR)) Bazofil granulociták esetében hasonló jelenség működik IgG ellenanyagokkal, és FcgReceptorral is. (természetesen a kis affinitású FcgR nem köt szabad ellenanyagot, hanem immunkomplexekkel reagál) pathogen expulsion (allergy symptoms)

FEATURES OF ANTIBODY-ANTIGEN INTERACTION - valency and avidity Valency: numbers of bonds between antigen and antibody Affinity: the strength of interaction between a specific antigen and one binding site of the antibody Avidity: „sum” of affinities of the binding sites of a given antibody Az Fc rész sajátoságainak – multimerizáció – mégis van jelentősége az a variábilis részhez köthető funkcióban (neutralizáció) is. A primer immunválasz során keletkező ellenanyagok gyakran kis affinitásúak (az affinitás érés még nem játszódott le), de az IgM multimer sajátsága ezt valamelyest kompenzálja aviditásban.

MAIN CHARACTERISTICS OF ANTIBODY ISOTYPES 1. valence free IgM pentamer (star shape) 2 2 2 IgA could be monomeric, dimeric, trimeric IgM could be monomeric, tetrameric, pentameric, hexameric 4 Antigen bound IgM (crab shape) 10

MAIN CHARACTERISTICS OF ANTIBODY ISOTYPES 2.

THE ROLE OF THE HINGE REGION

ANTIBODY PRODUCTION DURING THE PRIMARY AND THE SECONDARY IMMUNE RESPONSE alacsony magas antigénfüggő 100-1000x erősebb az első válasznál primer – elsődleges szekunder – másodlagos (tercier nincs, mert gyakorlatilag a szekunderrel azonos minden további immunizálás)

ANTIBODY PRODUCTION DURING THE PRIMARY AND THE SECONDARY IMMUNE RESPONSE level of antibodies secondary response against antigen A primary response against antigen A primary response against antigen B Ez csak egy példa a specificitásra – a második alkalommal egy másik független antigén együtt adva a korábbival. A másik antigén így is csak primer jellegű választ indukál napok Antigen A Antigen A and B

Isotype distribution in the body ANTIBODY ISOTYPE AND FUNCTION IgG Az izotípusok eloszlásáért többek közt „szállító” Fc receptorok felelősek Pink-blue heart represent – IgM and IgG predominance in the blood (circulation) Isotype distribution in the body

PRODUCTION OF IMMUNOGLOBULINS IgG IgM IgA AFTER BIRTH breast milk 100% (adult) 3 year 2 5 4 6 adult 9 1 month maternal IgG BEFORE BIRTH Magzatba anyai IgG kerül, és vam saját IgM termelése. Születés elötti IgM a természetes ellenanyagtermelés termékei (CD5+ B1 sejtek – gyakran evol.konzervatív struktúrájú antigének felismerése, DNS, HSP, hisztonok – a VDJ átrendeződésnél általában az első? Szekvenciák kódolják) Anyatej anyai IgA és IgM –jei védik a bélrendszer és a légutak felső szakaszait.

and transporting Fc receptors FcRn „neonatal” Fc receptor Placenta, endothelial and epithelial cells, etc. IgG transfer, IgG salvage pIgR poli Ig receptor Epithelial cells IgA, IgM transfer

Az IgG placental transport: neonatal Fcg receptor (FcRn) human FcgRn humán MHC Class I A placenta sejtek felveszik az anyai ellenanyagokat az anyai keringésből. Az endoszóma savas közegében az FcgRn megköti az ellenanyagokat (IgG specifikus), átszállítja a fötális oldalra, és az ott levő neutrálisabb pH hatására elengedi azokat. Egyes állatok esetében ez az előtejjel történik meg: Az újszülött bélrendszerében az FcγRn megköti a kolosztrumból származó anyai eredetü IgG-t (pH6.5). A receptor ezután transzcitózissal átszállítódik az epitéliumon és a megváltozott kémhatás hatására (pH 7.4) az IgG felszabadul és bekerül az újszülött állat keringésébe. (érdekesség- Az FcgRn és az MHC I molekula szerkezeti rokonságban áll.) Hasonló receptorok vannak az endotél sejtekben is. A pinocitózissal felvett ellenanyagokat ezek megkötik és visszaszállítják a felszínre – megmentik a lizoszómális lebontástól – növeli az ellenanyagok féléletidejét. Egérben nincs ilyen receptor, de bele lehet tenni – az ilyen egerekből sokkal több ellenanyag nyerhető ki (pl. diagnosztikai vagy kísérletes célokra) (the structure of FcRn and MHC I is related)

FcRn, FcγRn FcRn can transport IgG and albumin Újszülött rágcsálók bélrendszerében 100x magasabb az expressziója mint más szöveteken illetve az egyedfejlődés későbbi szakaszában. Náluk nincs placentális transzfer, hanem az anyatejből szerzi be a szervezetük az IgG-t. Rágcsálóknál az anyatejbe is ilyen receptorral kerülhet ki az anyai keringésből az IgG, főleg a laktáció elején (colostrum, fröccs tej). Azért közvetlenül a szülés után van, mert olyankor még nincsenek enzimek az újszülött rágcsáló bélrendszerében. Később nagyobb eséllyel emésztődik meg az IgG, mielőtt felvételre kerülne. Located in different cell types: endothelial cells epithelial cells (digestive and respiratory system) different immune cells eg. professional APC (cross presentation)

FcRn (FcγRn) function pinocytosis IgG is bound with high affinity in acidic conditions Az urogenitális traktusban, a rektumban és a felső légutak egy részében több az IgG, mint az IgA The Immunologic Functions of the Neonatal Fc Receptor for IgG J Clin Immunol (2013) 33 (Suppl 1):S9–S17; DOI 10.1007/s10875-012-9768-y Dendritikus sejtek cross-prezentációs folyamatában is lényeges szerepe van  Ag-Ab komplexek megmentése a lizoszómális lebontástól…… A rendszeres IVIG és friss fagyasztott szérum azért lehet hatékony egyes type II hiperszenzitivitásokban, mert a bejuttatott nagy mennyiségű IgG telíti az FcRn rendszert, és a „felesleges” nagy mennyiségű IgG ellenanyag az autoellenanyagokat is beleértve hamarabb lebomlik (kihígul) IgG can be released at the opposite side of the epithelia: transcytosis IgG can be transported to various sites of the body circulation  tissues tissues  epithelial surfaces (IgG is more abundant in the rectum and in some upper respiratory tract than IgA) maternal circulation  foetal circulation

Az FcRn salvages IgG from lysosomal degradation

SECRETORY IgA AND TRANSCYTOSIS ‘Stalk’ of the pIgR is degraded to release IgA containing part of the pIgR (the secretory component) J C S s J C S s J C S s MUC US J C S s J C S s IgA and pIgR are transported to the apical surface in vesicles J C S s Epithelial cell pIgR and IgA are internalised Polymeric Ig receptors are expressed on the basolateral surface of epithelial cells to capture IgA produced in the mucosa J C S s B B cells located in the submucosa produce dimeric IgA

multimeric IgM can exist without J chain, but not transported The cargo of pIgR (poli immunoglobulin receptor) Immunoglobulins with ”J-chain”: IgA, IgM (Do not confuse this J chain with the VJ/VDJ rearrangement’s J region!) multimeric IgM can exist without J chain, but not transported The destination epithelial surfaces, sercretums (saliva, breast milk)

SUMMARY OF THE EFFECTOR FUNCTIONS OF ANTIBODIES (next seminar)

MONOCLONAL ANTIBODIES POLYCLONAL ANTIBODIES and POLYCLONAL ANTIBODIES

POLYCLONAL ANTIBODY RESPONSE Ag Polyclonal antibody Immunserum Set of B-cells Ag Ag Activated B-cells Antibody-producing plasma-cells Antigen-specific antibodies

PRUDUCTION OF MONOCLONAL ANTIBODIES

MONOCLONAL ANTIBODIES products of one B lymphocyte clone homogeneous in antigen specificity, affinity, and isotype can be found in pathologic condition in humans (the product of a malignant cell clone) advantages against polyclonal antibodies: antibodies of a given specificity and isotype can be produced in high quantity and assured quality therapeutic usage of monoclonal ABs: anti-inflammatory antibodies (autoimmune diseases) tumor therapy

FEATURES OF POLYCLONAL AND MONOCLONAL ANTIBODIES Polyclonal antibody Monoclonal antibody (high affinity) Number of recognized antigen determinants several (frequent cross-reactions) mostly one Specificity polyspecific monospecific Affinity Varying (diverse antibodies) high Concentration of non-specific immunoglobulines low Cost of preparation Standardisability Impossible (or uneasy) easy Amount limited unlimited Applicability method-dependent excellent

POSSIBLE USE OF MONOCLONAL ANTIBODIES Identifying cell types Immunohistochemistry Characterization of lymphomas with CD (cluster of differentiation) markers Isolation of cells Isolation of CD34+ stem cells for autologous/allogeneic transplantation (from peripheral blood!) Blood group determination (with anti-A, anti-B, and anti-D monoclonals) Identification of cell surface and intracellular antigens Cell activation state Targeted chemotherapy CD20+ anti-B-cell monoclonals in non-Hodgkin lymphoma Prevention of organ rejection after transplantation

Monoclonal antibodies as drugs? Mouse monoclonal antibodies may elicit an immune response upon administration in human subjects. (see immunogenicity-determining factors!) How can we solve this problem?

EVOLUTION OF MONOCLONAL ANTIBODIES Mouse Human Humanized Chimeric Humanized antibodies are antibodies from non-human species whose protein sequences have been modified to increase their similarity to antibody variants produced naturally in humans.The process of "humanization" is usually applied to monoclonal antibodies developed for administration to humans (for example, antibodies developed as anti-cancer drugs). Humanization can be necessary when the process of developing a specific antibody involves generation in a non-human immune system (such as that in mice). The protein sequences of antibodies produced in this way are partially distinct from homologous antibodies occurring naturally in humans, and are therefore potentially immunogenic when administered to human patients Humanized antibodies are distinct from chimeric antibodies. The latter also have their protein sequences made more similar to human antibodies, but the fraction of foreign protein is larger than in humanized ones. Humanized antibodies are antibodies from non-human species whose protein sequences have been modified to increase their similarity to antibody variants produced naturally in humans.

Khimaira (görög betűkkel Χίμαιρα, latinosan Chimaera) nőstény szörnyalak a görög mitológiában. Szülei Tüphón és Ekhidna, testvérei a Hüdra, Kerberosz és a Szphinx és talán a nemeai oroszlán. Három feje van: elöl oroszlán, középen kecske, hátul sárkány (egyesek szerint kígyó), és lángokat okád. Úgy tartották, hogy megjelenése vihar, hajótörés, vulkánkitörés vagy más szerencsétlenség előjele. - Wikipédia -

humanized mouse monoclonal antibodies human monoclonal antibodies PASSZÍV IMMUNIZÁLÁS immunization mouse monoclonal antibodies Human immunoglobulin transgenic mouse humanized mouse monoclonal antibodies immunization human monoclonal antibodies PROTECTED SUBJECT serum antibody ENDANGERED SUBJECT This is a case of PASSIVE IMMUNIZATION Immune system is not activated prompt effect temporary protection/effect Immunoglobulin degradation

Passive immunization II. Type Usage Intramuscular HBV-Ig; Varicella-Zoster-Ig; Anti-D profilaxis Intravenal (IVIG) Humoral immunodeficiencies (Bruton agammaglobulinaemia; different deffects resulting hypogammaglobulinaemia) Anti-toxin, anti-viral antibodies (snake venom, Ebola) Natural passive immunization: maternal IgG placental transfer maternal IgA in breast milk

Passive immunization III. ”Antisera” / Policlonal antibodies Antivenom snakes, scorpions, spiders, dangerous toxic sea creatures Slow, gradial hyperimmunization of large animals (horses) Purification of the gamma globulin fraction from the sera of the hyperimmunized animals (Purified animal proteins (Ig) are milder immunogens than raw serum in the patients body. Repeated therapy could be possible)

MONOCLONALS AS DRUGS - Tumor therapy Monoclonals can be used for targeted chemotherapy of tumors. It is cell-type specific, but not specific to malignant cells! Immunsuppressive monoclonals Cell-type specific immunsuppression

MONOCLONALS IN TUMOR THERAPY „Naked MAb”, unconjugated antibody Anti-CD20 (rituximab – Mabthera/Rituxan, chimeric): B-cell Non-Hodgkin lymphoma Anti-CD52 (campath – Mabcampath, humanized): chronic lymphoid leukaemia Anti-ErbB2 (trastuzumab – Herceptin, humanized): breast cancer Anti-VEGF (bevacizumab – Avastin, humanized): colorectalis tu. (+ Lucentis!) Anti-EGFR (cetuximab – Erbitux, chimeric): colorectalis tu. (+ Vectibix, rekomb. humán!) 2. Conjugated antibody Anti-CD20 + yttrium-90 isotope (ibritumomab- Zevalin) Anti-CD20 + iodine-131 (tositumomab – Bexxar)

IMMUNSUPPRESSIVE ANTIBODIES Anti-TNF-α antibodies infliximab (Remicade): since 1998, chimeric adalimumab (Humira): since 2002, recombinant human Etanercept (Enbrel) – dimer fusion protein, soluble TNF-α receptor + IgG Fc-part Not a real monoclonal antibody. IgG Fc part  t1/2, transport See more in supplementary information Indications of anti-TNF-α therapy: Rheumatoid arthritis Spondylitis ankylopoetica (Bekhterev's disease) Psoriasis vulgaris, arthritis psoriatica Crohn’s disease, colitis ulcerosa (usually – still – not in the first line!)

CASE STUDY (MULTIPLE MYELOMA) In 1989, a 55-year-old housewife, who had been in good health her entire life, began to experience excessive fatigue. Her physician did not find abnormalities on physical examination. The blood sample revealed mild anemia; red blood cell count was 3.5 x 106 / ml (normal 4.2-5.0 x 106 / ml), white blood cell count was 3600 / ml (normal 5000 / ml). The sedimentation rate of her red blood cells was 32 mm / h (normal <20 mm / h). (Sedimentation is accelerated when fibrinogen or IgG content of the blood plasma is elevated.) The concentration of IgG was found to be 3790 mg / dl (normal 600 - 1500 mg / dl), that of IgA 14 mg / dl (normal 150 – 250 mg / dl) and that of IgM 53 mg / dl (normal 75 -150 mg / dl).

CASE STUDY (MULTIPLE MYELOMA) Electrophoresis of her serum revealed the presence of a monoclonal protein, which on further analysis was found to be IgG with lambda light chains. normal serum serum from the patient Radiographs of all of her bones did not show any abnormality. No treatment was advised.

CASE STUDY (MULTIPLE MYELOMA) In April 1991 her serum IgG was 4520 mg / dl, and in January 1992 it was 5100 mg / dl. By November 1992, her anemia had worsened and her red blood cell count had fallen to 3.0 x 106 / ml. At the same time her white blood count had fallen to 2600 / ml. In December 1992, she experienced the sudden onset of upper arm pain and headache. Radiographs of the skull and the left upper arm showed ‘punched out’ lesions in the bones.

CASE STUDY (MULTIPLE MYELOMA) She was treated with melphalan (methylphenylalanine mustard), corticosteroids, and irradiation. Her symptoms improved. In April 1993, further chemotherapy was given because of the persisting elevation of her serum IgG. The treatment reduced her serum IgG level from 8200 mg / dl to 6000 mg / dl. In February and in May 1995, she was found to have pneumonia. She was treated successfully with antibiotics. She recovered from this episode in the hospital and remained fully active. She required blood transfusion for her anemia and complained at times of bone pain. Her serum IgG was stable at 6200 mg / dl. Although she was in relative good health as our case history ended, her outlook for survival was very poor. Recently, bone marrow transplants have been used to cure patients with multiple myeloma. /Myeloma proteins have played an important part in the history of immunology. (Bence-Jones protein, subclasses of IgG, amino acid sequence of immuno- globulin molecule)/

Q&A The serum IgG from her was assumed to be monoclonal because it migrated as a tight band on electrophoresis in an agarose gel, and because it reacted with antibodies to lambda but not to kappa chains. What other evidence could be brought to bear to prove the monoclonality of this IgG? The IgG could also be shown to belong a single subclass of IgG, that is IgG1, IgG2, IgG3, or IgG4. Further more, it would be possible to show that a single variable-region gene was rearranged to form this IgG. She became anemic (low red blood cell count) and neutropenic (low white blood cell count). What was the cause of this? The proliferation of malignant plasma cells in the bone marrow crowded out blood cell precursors. This creates a limitation on space in the bone marrow.

As her disease progressed, she became susceptible to pyogenic infection; for example, she had pneumonia twice in a short period. What is the basis of her susceptibility to these infections? Although her serum IgG concentration is quite elevated, almost all the IgG is secreted by the myeloma cells and is monoclonal. In fact, she has very little normal polyclonal IgG and has been effectively rendered agammaglobulinemic by her disease. In addition, her white blood cell count is decreased and she has too few neutrophils (<1000 / ml) to ingest bacteria in the bloodstream and lungs effectively. A monoclonal immunoglobulin in the serum is called an M-component (‘M’ for myeloma). Is the presence of an M-component in serum diagnostic of multiple myeloma? No. M-component appear in the blood as people age. About 10% of healthy individuals in the ninth decade of live have M-component. This is called benign monoclonal gammopathy. Without bone lesions and presence of many malignant cells in the bone marrow, the diagnosis of multiple myeloma cannot be made. Some people have IgM M-components in their blood. This is due to another malignancy of plasma cells called Waldenström’s macroglobulinemia, which differs in many ways from multiple myeloma and is a more benign disease.

What happens with the B cells in myeloma multiplex? Healthy individual Myeloma multiplex

SUPPLEMENTARY INFORMATION

B CELL TUMORS

FcγRII mediated B cell feedback regulation Ag Ag B cell FcγRIIb B cell FcγRIIb Phosphorilated ITIM motifs on FcγRIIb recruits phosphatases to interfere signal transduction Inhibition of the signal transduction of B cell receptor

MONOCLONAL ANTIBODY NOMENCLATURE The nomenclature of monoclonal antibodies is a naming scheme for assigning generic, or nonproprietary names to a group of medicines called monoclonal antibodies. This scheme is used for the World Health Organization’s International Nonproprietary Names. Components of nomenclature: Example:Abciximabab- + -ci(r)- + -xi- + -mab, it is a chimeric monoclonal antibody used on the cardiovascular system

IMMUNSUPPRESSIVE ANTIBODIES 2. Muromonab-CD3 (OKT-3) egér IgG2a Against CD3 pan-T-cell antigen, after transplantation; It is rarely (or not) used nowadays (mouse protein!); ongoing trials in diabetes mellitus, with the humanized version Omalizumab (Xolair): Anti-IgE humanized IgG1k monoclonal Ind.: allergic asthma, Churg-Strauss sy. Daclizumab (Zenapax): anti-IL-2 receptor humanized antibody Ind.: transplantation basiliximab (Simulect): as daclizumab, but chimeric! efalizumab (Raptiva): anti-CD11a, humanized, used in psoriasis

MOLECULAR TARGETED DRUGS Name Type Target Indications Alemtuzumab (Mabcampath) Daclizumab (Zenapax) Basiliximab (Simulect) Rituximab (Rituxan/Mabthera) Trastuzumab (Herceptin) Gemtuzumab Ibritumomab (Y90) Edrecolomab Gefitinib Imatinib Monoclonal Ab, humanized Monoclonal IgG1, chimeric Monoclonal IgG1, humanized Monoclonal IgG4, humanized Calicheamicinnel konjugált Monoclonal IgG1, murine Monoclonal IgG2, murine EGFR-TKI KIT-TKI CD52 IL-2 R CD20 HER2/neu CD33 EpCAM EGFR TK TK CLL, CML transplantation Lymphoma Breast cancer, NSC lung cancer leukemia lymphoma CRC NSCLC GIST, CML

FURTHER POSSIBILITIES WITH MONOCLONALS Radioimmunotherapy As Zevalin, Bexxar – monoclonal + isotope Antibody-directed enzyme prodrug therapy (ADEPT) An enzyme is linked to the antibody, and the enzyme will make citotoxic drug from the later administered prodrug Immunoliposomes Targeting nucleotides or drugs in liposomes, linked to an antibody Non-immunological targets as abciximab (ReoPro): inhibition of thrombocyte-aggregation

Chimeric Antigen Receptors (CAR) Effector cytotoxic or cytokin producing helper T cells transfected with engineered antigen receptor construct. first generation second generation third generation chimeric antigen receptors (CARs) linker variable domains of ”tumour antigen” specific antibody VH VL antibody hinge region Marcela V. Maus, Stephan A. Grupp, David L. Porter, and Carl H. June Antibody-modified T cells: CARs take the front seat for hematologic malignancies BLOOD, 24 APRIL 2014 x VOLUME 123, NUMBER 17 signal transduction domains directly linked to the receptor

TRUCK (T cells redirected for universal cytokine killing) CAR T cells could also be co-transfected with inducible pro-inflammatory, cytotoxic, immunstimulatory cytokine gene constructs: TRUCK (T cells redirected for universal cytokine killing) Markus Chmielewski, Andreas A. Hombach, Hinrich Abken: Of CARs and TRUCKs: chimeric antigen receptor (CAR) T cells engineered with an inducible cytokine to modulate the tumor stroma Immunological Reviews 2014, Vol. 257: 83–90

Gianpietro Dotti, Stephen Gottschalk, Barbara Savoldo, Malcolm K Gianpietro Dotti, Stephen Gottschalk, Barbara Savoldo, Malcolm K. Brenner: Design and development of therapies using chimeric antigen receptor-expressing T cells Immunological Reviews 2014, Vol. 257: 107–126