INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent...
Transcript of INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent...
![Page 1: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/1.jpg)
INMUNOLOGIA MOLECULAR
![Page 2: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/2.jpg)
HUMORAL CELULAR
INMUNIDAD IgM MacrófagosNATURAL Células NK
(sin memoria) Neutrófilos
INMUNIDAD IgG Linfocitos CD4+ ADAPTATIVA (helper)(con memoria) Linfocitos CD8+ (CTL)
JM/03Medicina Molecular
![Page 3: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/3.jpg)
Pathways of differentiation of a pluripotenthematopoietic stem cell of the bone marrow
136380
differentiate in the thymus
differentiate in the bone marrow
Stemcells
![Page 4: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/4.jpg)
ANTIGENOS
Moléculas reconocidas por receptores en células B (B cellreceptor) o T (T cell receptor).
Hapteno : antígeno que para ser reconocido debe ser unido a un
carrier (hidratos de carbono)
![Page 5: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/5.jpg)
How many environmental antigens are we exposed to
during our lifetime?
The immune system responds to hundreds of thousands of foreign antigens introduced from the environment
![Page 6: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/6.jpg)
Examples of antigen, immunogen and epitopes
Antigen
Epitope A Epitope B
Immunogen
Epitope B
Antibody A Antibody B
Antibody B
![Page 7: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/7.jpg)
![Page 8: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/8.jpg)
ANTICUERPOS
![Page 9: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/9.jpg)
![Page 10: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/10.jpg)
![Page 11: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/11.jpg)
![Page 12: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/12.jpg)
![Page 13: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/13.jpg)
RECEPTORES ANTIGENICOS
• B cell receptor (linfocitos B) = reconoce Ag nativo
• T cell receptor (linfocitos T) = reconoce Ag procesado
JM/00Medicina Molecular
![Page 14: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/14.jpg)
ANTIGENO CON MULTIPLES EPITOPES
SUERO CON MULTIPLES ANTICUERPOS
![Page 15: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/15.jpg)
![Page 16: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/16.jpg)
![Page 17: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/17.jpg)
APCB Ab
CD4
CD8
Ag
Ag Processing
HelpHelp
1. 2.
The Immune Response:
![Page 18: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/18.jpg)
LINFOCITO CD4 ACTIVADO
![Page 19: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/19.jpg)
Role of T-Helper cells in antibody formation.
![Page 20: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/20.jpg)
LINFOCITO B EN REPOSO
![Page 21: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/21.jpg)
LINFOBLASTOS B
![Page 22: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/22.jpg)
![Page 23: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/23.jpg)
Lymph node
![Page 24: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/24.jpg)
Germinal center in lymph node
![Page 25: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/25.jpg)
PLASMOCITO
![Page 26: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/26.jpg)
PLASMOCITO APOPTOTICO
![Page 27: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/27.jpg)
Primary and secondary antibody responses.
136384
![Page 28: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/28.jpg)
¿Cómo se produce la gran diversidad de anticuerpos?
![Page 29: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/29.jpg)
Susumu TonegawaPremio Nobel 1987
![Page 30: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/30.jpg)
Producción de cadena κ
![Page 31: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/31.jpg)
GENES CODIFICANTES PARA INMUNOGLOBULINAS
• Cluster IGH: cadena H; crom.14;V,D,J,C.
• Cluster IGK: cadena K; crom.2; V,J,C.
• Cluster IGL(λ): cadena λ; crom. 22; V,J,C.
![Page 32: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/32.jpg)
![Page 33: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/33.jpg)
![Page 34: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/34.jpg)
ANTICUERPOS MONOCLONALES
![Page 35: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/35.jpg)
b. 1927(in Bahia Blanca, Argentina)d. 2002
b. 1946d. 1995
b. 1911d. 1994
MRC Laboratory of Molecular BiologyCambridge, United Kingdom
Basel Institute for ImmunologyBasel, Switzerland
Basel Institute for ImmunologyBasel, Switzerland
Argentina and United KingdomFederal Republic of GermanyDenmark
1/3 of the prize1/3 of the prize1/3 of the prize
César MilsteinGeorges J.F. KöhlerNiels K. Jerne
Find a Laureate:
![Page 36: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/36.jpg)
• antibodies produced by differentiated B-cells
• recombination results in B-cell lineages producing distinct antibodies
• serum contains antibodies representing many lineages
• difficult to isolate B-cells of defined specificity
• cannot grow B-cells in culture
Kohler and Milstein (1975)
• fused B-cells with myelomacells stable hybridoma
• hybridoma secretes monoclonal antibody
![Page 37: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/37.jpg)
• immunized mouse making desired antibodies
• boost 3-4 days before fusion• remove spleen and harvest
cells• mix with myeloma cells in
presence of fusogenic agent
Balb/c Mouse
MOPC 21 (tumor)
P3K (cell line)
P3-X63Ag8 (HGPRT-)
X63-Ag8.653 (IgG-)
![Page 38: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/38.jpg)
HGPRT = hypoxanthine-guanine phosphoribosyl transferaseDHFR = dihydrofolate reductase
HAT Medium:• Hypoxanthine
(purine salvage)• Aminopterin
(DHFR inhibitor)• Thymidine
(pyrimidine salvage)
Nucleotide Metabolism:
• salvage and de novo pathways
• HGPRT essential for purine salvage
• DHFR essential for de novo synthesis
![Page 39: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/39.jpg)
Selection in HAT Medium
Human mAbs produced by trans-forming lymphocytes with EBV
![Page 40: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/40.jpg)
Cloning Methods•soft agar• limiting dilution
Screening/Cloning
![Page 41: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/41.jpg)
• produce mAbs in vitro or with ascites
• harvest culture media (supernatant)– in vitro material is less
concentrated and contains bovine serum
• ascites are tumors grown in peritoneal cavity– ascites fluid and sera
contain high [mAb]– minor contamination
with mouse IgG
Production
![Page 42: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/42.jpg)
HAMA ("human anti-mouse antibodies"). HAMA ("human anti-mouse antibodies").
![Page 43: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/43.jpg)
Mouse monoclonal antibodies have been genetically engineered to replace all of the antibody molecule with human counterparts except the hypervariableregions directly involved with antigen binding.Humanized monoclonal antibodies are
currently be tested in human clinical trials.
Humanized Monoclonal Antibodies
![Page 44: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/44.jpg)
![Page 45: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/45.jpg)
RECEPTORES ANTIGENICOS
• T cell receptor (linfocitos T) = reconoce Agprocesado
• B cell receptor (linfocitos B) = reconoce Agnativo
![Page 46: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/46.jpg)
![Page 47: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/47.jpg)
MECANISMO DE ACCION DE LOS ANTICUERPOS
Via ComplementoADCC (antibody-dependent-
cellular-cytotoxicity
![Page 48: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/48.jpg)
![Page 49: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/49.jpg)
![Page 50: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/50.jpg)
• COMPLEMENTO
![Page 51: INMUNOLOGIA MOLECULAR I.ppt [Sólo lectura] · Pathways of differentiation of a pluripotent hematopoietic stem cell of the bone marrow 136380 differentiate in the thymus differentiate](https://reader030.fdocuments.in/reader030/viewer/2022040701/5d59a61f88c993c4088b63f4/html5/thumbnails/51.jpg)
MAC (complejo ataque complemento)