Exam 26.01.24 08.15-08-30 The two were really nice and they let me choose in which language I want to take the exam.
Halin: Vaccine conjugates - What's the principle behind vaccines? Immunization - Which type of immune response? BC, CD4 (I kept it quite simple, but I think she likes if you go through all the details, incl. presentation on MHC, etc.) - In cancer would another cell type be preffered, which? CD8 - Why aren't they prefferably activated? (She helped me to get to uptake by DCs and MHC II presentation) - Concept of conjugate vaccines? Explained HiB vaccine, linked recognition, mentioned act. of various BCs by presentation of the same peptide to the same CD4 TC
Neri: Insulin - Tell about history and insuline. oldest recombinant protein, already isolated over 100 years ago from bovine pankreas, alpha-/beta-chain, difference between mono-/di-/hexamer and started talking about modifications (short: lispro (Lys/Pro switched, no di-/hexamer formation; long: glargine (2 Arg: incr. pI, red. solubility) determir (myristolyation: COOH + alkyl chain to mimic fatty acid and enables binding to albumin -> incr. t1/2) - why has albumin long t1/2? started to talk about FcRn because i couldn't remember anything else (+ size and thereby decr. filtration in kidney) - Recombinant production of insulin not easy because of two chains. How would you produce? E. coli, two methods: 2 chains + SS-bond in periplasm or 1 chain & cutting - Why E.coli? Insulin is simple (no glycosylation etc.) and explained E. coli advantages - He ended with: "Yes, is simple, fast and cheap" and nodded
The exam ended quite quick and it felt more like a discussion than really being examined. Whatch out for the following points: Halin seems to like hearing all the details, especially if you're talking about the incuction of the immune response. Go through them step by step. Neri sometimes interrupted me when i was searching for specific details like the name of e.g. an amino acid, he then quickly gave the answer and told me that it's also not that important and he'd likes to hear the concept. So try to be concrete, e.g. his last statement about E. coli "simple, fast and cheap".
Immunoglobulins Was sind immunoglobulins? Welche gibt es und wo kommen sie vor? wofür wichtig? Wie kommt IgG über plazenta? Wieso IgG nach 6 monate in baby tief? Wenn keine antibodies welche infektionen? ...
Engineering Was kann man engineeren? Bezogen auf Ab: Wie kann man bindungsaffinität erhöhen? mutation in cdr halbwertszeit verlängern? (nicht Fc, da schon Fc) kann man Ab glykosilieren? Gibt medikament (Markenname), wo das gemacht wurde gegen CD20? wieso wurde das gemacht. stichwort bindungsaffinität. ADCC --> Was ist ADCC --> aktiviert NK
-Alles zu Morbus Crohn erzählen - dann wollte sie vor allem wissen was Th1 und Th17 machen und deshalb was neutrophile machen. - Es können granulom entstehen --> was sind granulom?
Neri: Clearance of therapeutic proteins
-Wie sind therapeutic proteins cleared? - wie gross darf ein Molekül sein, damit es von der Niere ausgeschieden wird? - über pegylation geredet, um half-life zu verlängern - FcRn recycling genauer erklärt/ gesprochen - was wird bei Insulin zb detemir gemacht damit längere halbwertszeit?
T regulatory cells What do they do? --> help regulate immune responses, suppress overreactive T-cells, help tumors Where are they formed? --> during development in Thymus or in sec. lymphoid structure. How are they formed/What is needed? --> TCR-MHC binding, Co-stimulation ratio between inhibitory and stimulatory factors lead to TH cells or Treg cells. If the stimulation isn’t strong enough -> Treg
Production of human monoclonal antibodies How are they produced? --> transgenic mouse with fully human immunoglobulin locus, phage display Areas in which mAbs can be used. (Immunosuppression, tumors, inflammatory diseases, ocular therapy (VEGF), Hypercholesterinemia (PCSK9))
Halin: Morbus Crohn - could tell everything I knew about the disease - about NOD2 / pattern recognition receptors in general: what do they recognize? - what happens if a bacterium reaches the submucosa? - why are TH1 / TH17 responses mounted? what cytokines are released? what are the effects? - do you know a disease where IL-17 plays a role in the disease pathology? - what happens in the presence of non- invasive bacteria?
Neri: Clearance of therapeutic proteins - how are small molecules / big molecules excreted? (small: kidney, big: liver) - why do antibodies have a long half-life? - explanation of FcRn recycling - do you know of any therapeutic proteins that have a short half-life? - Abciximab: Fab-fragment hat binds GPIIb/IIIa repector on platelets and inhibits platelet aggregation - why does plattelet aggregation returns to normal only after 4-5d? (due to its high affinity)
26.01.24 11:45-12:05 Halin: Cancer immunotherapy -What is it, give a definition, explain it in your own words - Explain and elaborate on different methods that can be used (I mentioned CTLA-4/PD-1 inhibitors and T-VEC and had to explain all three in detail) Halin wanted to know details, such as what part binds to exactly what, so don't ignore Immuno I.
Neri: Small molecules vs antibodies -What are the differences, name the pros and cons of both -Can both be immunogenic? Why, why not? The Neri part was cut a bit short because Halin took up too much time in the first part
Autoimmunity: what is autoimmunity? what leads to autoimmune disease? question about self tolerance? breaking self tolerance? Example t cell mediated autoimmune disease?
bioequivalence: What is it? why important and when is it important? talked about MS, Chromatography...
Halin The topic is “Major histocompatibility and minor histocompatibility complex” Q:She asked me to talk about it A:I said that each individual has different MHC and this will lead to the immune response after transplant Q:She than asked me what kinds of MHC there are A:I didn’t get it(I told her there are different kinds of HLA genes but I can’t list all of them)then she said it’s MHC1&2 lol Q:Next she asked me which MHC induce which cell(CD4&CD8)and which MHC express by which cell(MHC1 express on all the cells but not only APC）
A:Then I talked about what minor histocompatibility is I said each individual has different amino acid sequences which lead to slight differences among each individual’s protein sequence And thus in transplantation, even though they do the MHC matching in advance, there might happen immune response due to the recognition of those proteins with different amino acid sequences (She also asked one more question at the end but I didn’t really get it and forgot what is it now lol
Neri He asked me to talk about any antibodies for cancer therapy And I talked about antibody drug conjugate(ADC) He asked about mechanisms& And also what the target I’m a bit confused about the target part(I said tumor&tumor associated antigen, it’s correct but not what he want)in the beginning But then he guided me a bit I then realized he want me to talk about specific tumor associated/specific antigen(I picked HER2&BCR-AI, respectively)
Halin: Atopie Was ist atopie? Wie funktioniert allergie im allgemeinen? Was passiert während der sensibilisierungsphase? Warum juckt es bei allergie? -> histamine Was sind Eigenschaften von allergenen? Wieso nicht jeder allergisch? (sie wollte da auf Tregs hinaus) Es gibt ja genetische einflüsse für allergien, was hat die mutation in dem gen für IL4 für auswirkungen?
Neri: Antikörper welche cytokine beinflussen Nenne einige, ihr Target und wie sie funktionieren -> Humira (TNF- blocker, blockieren nicht nu sonder bilden auch komplexe die gecleared werden können) Dann was kann man denn noch weiter downstream vom TNF targeten? -> ich hab ihm dann von IL-17 und IL-23 blockern für Psoriasis erzählt. Ok welche Zellen sind denn am ende von dem Pathway den IL-17/IL-23 auslösen? -> neutrophile Dann es gibt ja statt humira auch noch andere neue medis die small molecules sind gegen RA, wie wirken die denn? -> oral JAK/STAT inhibitors, blockieren Phsophorlierung und blockieren somit die Interaktion vom Cytokin am cytokin rezeptor
Der Halin Teil braucht die meiste Zeit der Prüfung (ca. 2/3)
30.1.24 10:30-10:45 Halin: XLA What is it?: X linked immunodeficiency, btk mutation --> no mature b cells, no Abs For which type of pathogen could it be a problem?: Extracellular bacteria (Opsonization impaired) How do the other types of pathogens get handled?: Intracellular bacteria--> Macrophages (phagosome, lysosome) --> Th1 cells How can you measure it?: ELISA or FACS Is it severe?: No you can live with it How would you treat it?: i.v. IgG
Neri: Blood products Started talking about erithropoietin, but he said that's not blood derived --> Blood coagulation products: Factor VII: Hemophilia A Factor IX: Hemophilia B Hemlibra: bispecific Ab that links factor IX and X (imitates VIIIa function) Why would you want an Ab that imitates function instead of actual product?: I didn't know, he asked if it would be immunogenic?: Yes bc the patients don't have the factor the immune system would see it as foreign, even though it's a natural product He mentioned there's also Reopro, what is it?: GpIIa/IIb, for platelet production Does it stimulate or block platelet production? Why?: Blocks it, used for coronary interventions How long do Fab fragments stay in blood?: I didn't know, he said 30 min But effect of reopro lasts longer, why?: because it has a very high affinity--> once it binds, it stays long
They were very nice and the 15 min went by quickly
Halin: What’s a vaccine and what is it used for? To prevent infection ect. Herd immunity, for which vaccines did it not work? – HIV, tetanus I had to explain what an adjuvant was. – put the immune system in a state of alert and upregulates MHC expression and production of co-stimulatory molecules like B7.
Neri: What is a bispecific AB? Acts like a bridge between T cell and tumor cell, is a MHC-independent killing of the tumor cell, T cell releases perforins and granzymes to kill the tumor ect How is it produces? – knobs to holes (like a puzzle) What’s the problem with it? – Accessibility, targeting
Halin: Transplant rejection - What do you know about transplant rejection? -> explained blood group etc. - What does IgM do? (in the context of blood group) -> complement activation - End-point effect of complement activation? (in the context of blood group) -> opsonisation of erythrocytes by macrophages, pore formation of erythrocytes - What about MHC? -> polygenic, polymorph, direct & indirect rejection of graft - From which family members would you like to receive an organ transplant from? -> from siblings, 25% chance of having same haplotypes - If the MHC and blood group match, is it possible that the transplant will no longer be rejected? -> no, minor histocompatibility antigens different (due to SNPs)
Neri: Blood-derived therapeutic proteins - Name one conventional therapeutic protein that is still used (e.g. derived from blood serum) -> antibody - Why/For which patients? -> people with immunodeficiencies (e.g. XLA) - Name one blood coagulation drug -> Hemlibra, explained function etc. - Why not just inject recombinant factor VIII? -> immunogenicity - In the lecture: saw a video about Hemlibra in clinical trial. What was measured/How was the efficacy of Hemlibra determined? -> measured bleeding of patients - What is Reopro? -> fab fragment, block GPIIb/GPIIIa receptor on platelets - Half-life of fab fragment? -> I guessed 1h - How is fab cleared? -> through kidney - Why use fab instead of antibody? -> don't want aggregation of platelets for a long time - Name 3 events that may lead to blood coagulation (Virchow's triad) -> stasis of blood flow, endothelial injury, hypercoagulability
Halin: ABO and Rhesus System - Tell me what these are (Just talked about them; mention every detail) - Detection of ABO System (Agglutination but also explain FACS..)
Neri: GMP Production process of Humira: tell me the whole process from cell banks until the filling of the product (Bit confused what he wanted to hear; talked randomly about things that came into my mind.) -> Things he wanted to hear were: Control of viral contaminants (name one test), Purification methods: SDS Page, SEC and before filling filtration (Nano and sterile)
Halin: T-Cell mediated Autoimmunity (I directed the follow up questions by using different words) - Definition of Autoimmunity: Immune reaction towards self-antigen e.g. T-Cell mediated Autoimmunity MS, where T-Cell induce immune response towards MBP (Microgliacells) - Self-tolerance: peripheral anergy, Central tolerange, antigen segregation, B-Cell tolerance, Treg, differentiation and apoptosis - B-Cell maturation: bone marror proliferation -> pre-B-Cell -> peripheral maturation - Bone marrow: heavy chain rearrangement - Central Tolerance: AIRE - selfantigen production for neg. selection, failed neg. selection are eliminated or become T-reg
Neri: Small molecule vs Ab (directed by neri) - Difference: size, extravation-ability, half-live - Size of Ab and small molecule - Why is small molecule more toxic than Ab (he wanted to hear that small molecules are intracellular and extracellular binding, and Ab only extracellular and target specific)
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