- Spleen tyrosine kinase (Syk) is a novel pharmaceutical target in the treatment of allergic, autoimmune and neoplastic diseases. Previous studies have shown that Syk signaling plays a key role in regulating the lymphohematopoietic system. These observations prompted us to investigate whether Syk’s inhibition promoted immunotoxicity.
- In a series of studies, rats were given orally R406 at doses up to and including 100 mg / kg / day (or its prodrug R788 at doses up to and including 100 mg / kg / day, reduced to 50 mg / day). ). kg / day for women after passing MTD), a strong Syk inhibitor, twice daily for 28 days.
- Oltre alle valutazioni tossicologiche standard, sono stati eseguiti test di immunofenotipizzazione citometrica a flusso e di risposta immunitaria umorale misurando i livelli di anticorpi IgM e IgG anti – KLH . Altri studi di immunotossicità includevano tre modelli di resistenza dell’ospite in topi Balb/c femmine per determinare ulteriormente gli effetti di R406 sull’immunità innata e acquisita.
- Dopo il trattamento con R406, negli studi sui ratti sono stati osservati gli effetti immunomodulatori attesi (ad es. diminuzione del peso del timo e della milza, perdita di cellule del midollo osseo e diminuzione della conta dei linfociti, inclusi i linfociti T e B). Questi cambiamenti si sono generalmente risolti dopo un periodo di recupero di 14 giorni senza trattamento . KLH challenge nei ratti non ha mostrato effetti avversi sulla risposta IgG o IgM . R788/406, somministrato per via orale a dosi fino a 80 mg/kg/die inclusi per 28 giorni, non ha influenzato la clearance batterica o virale nei modelli di resistenza murina rispettivamente di Listeria, streptococco o ospiti dell’influenza.
- Ciò era correlato a precedenti test di funzionalità dei macrofagi e dei neutrofili in vitro (valutazione di ioni migratori, fagocitosi, scoppio ossidativo e attività battericida) che hanno mostrato che l’ R406 non ha influenzato negativamente la funzione dei macrofagi o dei neutrofili nelle risposte immunitarie innate. Presi insieme, questi risultati mostrano che l’ R406 ha un’immunotossicità funzionale minima nonostante i suoi effetti linfocitopenici, suggerendo che l’inibizione di Syk potrebbe non portare a effetti avversi inaccettabili basati sui meccanismi.
Valutazione dell’immunotossicità per il nuovo inibitore della tirosin-chinasi della milza R406.
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- Cdna 8Spleen tirosin chinasi (Syk) è un nuovo target farmaceutico nel trattamento delle malattie allergiche, autoimmuni e cancerose. Precedenti studi hanno dimostrato che la segnalazione Syk svolge un ruolo chiave nella regolazione del sistema linfoematopoietico.
- Queste osservazioni ci hanno spinto a indagare se l’inibizione di Syk promuovesse l’immunotossicità. In una serie di studi , ai ratti è stato somministrato per via orale R406 a dosi fino a 100 mg/kg/die inclusi (o il suo profarmaco R788 a dosi fino a 100 mg/kg/die inclusi, ridotti a 50 mg/die). kg/die per le donne dopo aver superato la MTD), un forte inibitore Syk, due volte al giorno per 28 giorni.
- In addition to standard toxicological assessments , flow cytometric immunophenotyping and humoral immune response tests were performed by measuring levels of anti – KLH IgM and IgG antibodies . Other immunotoxicity studies included three host resistance models in female Balb / c mice to further determine the effects of R406 on innate and acquired immunity.
- After treatment with R406, the expected immunomodulatory effects (e.g. decreased thymus and spleen weight, loss of bone marrow cells and decreased lymphocyte counts, including T and B lymphocytes) were observed in rat studies. .
- These changes generally resolved during the 14-day treatment-free recovery period. KLH challenge in rats showed no adverse effects on IgG or IgM response . R788 / 406, administered orally at doses up to and including 80 mg / kg / day for 28 days, did not affect bacterial or viral clearance in mouse resistance models of Listeria, streptococcus or influenza hosts, respectively.
- This was related to previous in vitro macrophage and neutrophil function tests (evaluation of migratory ions, phagocytosis, oxidative burst and bactericidal activity) which showed that R406 did not adversely affect macrophage or neutrophil function in immune responses. innate.
- Taken together, these results show that R406 has minimal functional immunotoxicity despite its lymphocytopenic effects, suggesting that Syk inhibition may not lead to unacceptable mechanism-based adverse effects.
To test whether IL-10 acts as an autocrine or paracrine growth factor for Ly-1 B cells, we treated mice continuously from birth to 8 weeks of age with a rat monoclonal IgM antibody that specifically neutralizes IL-10. murine. Mice treated in this manner were deprived of peritoneal Ly-1 B cells, contained significantly reduced serum levels of immunoglobulin M, and were unable to generate significant in vivo antibody responses to intraperitoneal injection of alpha 1,3-dextran or phosphorylcholine, antigens , for which specific B cells are found in the subset of Ly-1 B cells.
In contrast, spleen B cells from conventional anti -IL-10-treated mice were normal with respect to total number, phenotype, and in vitro response to B cell mitogens and trinitrophenyl hemocyanin antigen (TNP- KLH ) from thymus-dependent keyhole patella. ).
The mechanism of Ly-1 B cell depletion appears to be related to increased levels of endogenous interferon gamma (IFN-gamma) in mice treated with anti-IL-10 because they co-administered anti-IFN-gamma sub anti – IL. – 10 neutralizing antibodies . alli restored the number of Ly-1 B cells residing in the peritoneum of these mice.
In contrast, spleen B cells from conventional anti -IL-10-treated mice were normal with respect to total number, phenotype, and in vitro response to B cell mitogens and trinitrophenyl hemocyanin antigen (TNP- KLH ) from thymus-dependent keyhole patella. ).
The mechanism of Ly-1 B cell depletion appears to be related to increased levels of endogenous interferon gamma (IFN-gamma) in mice treated with anti-IL-10 because they co-administered anti-IFN-gamma sub anti – IL. – 10 neutralizing antibodies . alli restored the number of Ly-1 B cells residing in the peritoneum of these mice. These results suggest that IL-10 is a growth regulator of Ly-1 B cells and identify a procedure for specific depletion of Ly-1 B cells, thus allowing further evaluation of the role of these cells in the immune system.
Anti-Calnexin antibody (KLH)
STJ97891
100 µl
234 EUR
Anti-Calregulin antibody (KLH)
STJ97892
100 µl
234 EUR
Anti-cAMP antibody (KLH)
STJ97893
100 µl
234 EUR
Anti-CD20 antibody (KLH)
STJ97914
100 µl
234 EUR
Anti-Clenbuterol antibody (KLH)
STJ97960
100 µl
234 EUR
Anti-Flag antibody (KLH)
STJ98077
100 µl
234 EUR
Anti-MTAP Antibody (KLH)
STJ503536
100 µg
586 EUR
Mouse Keyhole Limpet Hemocyanin (KLH IgM) ELISA Kit
abx053480-96tests
96 tests
668 EUR
Human Anti-Meningococcal Group A/C/W/Y Oligosaccharides IgM (Combo) ELISA Kit, 96 tests, Quantitative
600-884-IGM
1 Kit
1017 EUR
Human Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-145-KLM
1 Kit
773 EUR
Mouse Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-155-VAM
1 Kit
712 EUR
Monkey Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-180-KLM
1 Kit
773 EUR
Canine Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-190-KLM
1 Kit
773 EUR
Rat(anti-CMV IgM)
QY-E11747
96T
374 EUR
Rabbit anti Rat IgM
40C-CB1264
2 mg
285 EUR
Goat anti Rat IgM
40C-CB1271
1 mg
363 EUR
Rat anti Mouse IgM
41R-1116
100 ug
265 EUR
Mouse anti Rat IgM
41R-1122
100 ug
265 EUR
Mouse anti Rat IgM
41R-1123
100 ug
265 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-b-48T
48T
456 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-b-96T
96T
590 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Ch-48T
48T
426 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Ch-96T
96T
549 EUR
Human Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Hu-48T
48T
404 EUR
Human Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Hu-96T
96T
518 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Mu-48T
48T
411 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Mu-96T
96T
528 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Rb-48T
48T
508 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Rb-96T
96T
661 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-b-48Tests
48 Tests
473 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-b-96Tests
96 Tests
654 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Ch-48Tests
48 Tests
437 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Ch-96Tests
96 Tests
603 EUR
Human Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Hu-48Tests
48 Tests
411 EUR
Human Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Hu-96Tests
96 Tests
565 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Mu-48Tests
48 Tests
419 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Mu-96Tests
96 Tests
578 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Rb-48Tests
48 Tests
534 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Rb-96Tests
96 Tests
742 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RD-IgM-b-48Tests
48 Tests
453 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RD-IgM-b-96Tests
96 Tests
625 EUR
Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit
CSB-E13375m-24T
1 plate of 24 wells
165 EUR
Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit
1-CSB-E13375m
Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit
CSB-E16534h-24T
1 plate of 24 wells
165 EUR
Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit
1-CSB-E16534h
Anti-Ki-67 antibody (KLH)
STJ98194
100 µl
234 EUR
Anti-Ki-67 antibody (KLH)
STJ98195
100 µl
234 EUR
Anti-PARP-1 antibody (KLH)
STJ98308
100 µl
234 EUR
Anti-Actin beta antibody (KLH)
STJ97814
100 µl
234 EUR
Anti-Bcl-2 antibody (KLH)
STJ97860
100 µl
234 EUR
Anti-E alpha antibody (KLH)
STJ98053
100 µl
234 EUR
Anti-Myc-tag antibody (KLH)
STJ90105
200 µl
197 EUR
Anti-His-tag antibody (KLH)
STJ90106
200 µl
197 EUR
Anti-HA-tag antibody (KLH)
STJ90107
200 µl
197 EUR
Anti-FLAG-tag antibody (KLH)
STJ90108
200 µl
197 EUR
Lyophilized Exosome Standards
ExoA375
100 μg
303.5 EUR
Lyophilized Exosome Standards
ExoHT29
100 μg
303.5 EUR
Lyophilized Exosome Standards
ExoMCF7
100 μg
303.5 EUR
Lyophilized Exosome Standards
ExoPC3
100 μg
303.5 EUR
Lyophilized Exosome Standards
ExoPLASMA
100 μg
303.5 EUR
Lyophilized Exosome Standards
ExoUrine
100 μg
303.5 EUR
Human Anti-Keyhole limpet hemocyanin (KLH) IgG+IgM ELISA Kit, 96 tests, Quantitative
700-135-KGM
1 kit
773 EUR
Bovine/Cow Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-205-KLM
1 Kit
712 EUR
Rat KLH ELISA Kit
ERK0075
96Tests
521 EUR
Goat anti-Rat IgM Antibody
abx401442-05mg
0.5 mg
370 EUR
Goat Anti-Rat IgM-AP
LF-SA8011AP
1.0 ml
353 EUR
Goat Anti-Rat IgM-BIOT
LF-SA8011B
1.0 mg
312 EUR
Goat Anti-Rat IgM-FITC
LF-SA8011F
1.0 mg
274 EUR
Goat Anti-Rat IgM-HRP
LF-SA8011H
1.0 ml
280 EUR
Goat Anti-Rat IgM-PE
LF-SA8011P
0.5 mg
437 EUR
Goat Anti-Rat IgM-TXRD
LF-SA8011T
1.0 mg
245 EUR
Goat Anti-Rat IgM-TRITC
LF-SA8011TR
1.0 mg
266 EUR
Goat Anti-Rat IgM-UNLB
LF-SA8011U
1.0 mg
261 EUR
Rabbit anti Rat IgM (FITC)
43C-CB1265
1.5 mg
289 EUR
Rabbit anti Rat IgM (rhodamine)
43C-CB1266
1.5 mg
303 EUR
Rabbit anti Rat IgM (biotin)
43C-CB1268
1.5 mg
368 EUR
Rabbit anti Rat IgM (HRP)
43C-CB1269
1.5 mg
399 EUR
Goat anti Rat IgM (FITC)
43C-CB1272
1 mg
364 EUR
Goat anti Rat IgM (rhodamine)
43C-CB1273
1 mg
402 EUR
Goat anti Rat IgM (biotin)
43C-CB1275
1 mg
389 EUR
Goat anti Rat IgM (HRP)
43C-CB1276
1 mg
445 EUR
Mouse anti Rat IgM (HRP)
43R-1632
100 ug
282 EUR
Mouse anti Rat IgM (HRP)
43R-1633
100 ug
282 EUR
Polyclonal Goat anti-GST α-form
GST-ANTI-1
50 uL
280 EUR
These results suggest that IL-10 is a growth regulator of Ly-1 B cells and identify a procedure for specific depletion of Ly-1 B cells, thus allowing further evaluation of the role of these cells in the immune system.