i.v. injection of human recombinant TNF-alpha

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ltrbrown
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i.v. injection of human recombinant TNF-alpha

I am hoping that someone may have some experience with i.v. injection of human recombinant TNF-alpha. I am going to test the effects of this protein on the growth potential cell lines in Nude mice. My research of the literature shows doses that range from 4 ug to 10 ug ("max tolerated dose") per mouse, by i.v. injection. Could anyone advise about these number and suggest possible symptoms that indicate toxicity (weight loss and others?). Most people did a single injection and followed tumor growth for up to 12 days, which indicates that TNF-alpha is pretty stable. My Supervisor would like me to dose at 8ug per mouse per week for 3 weeks, but I feel this may be excessive.

marcus muench
marcus muench's picture
 TNF hits mice (and humans)

 TNF hits mice (and humans) very hard.  It is one of the few cytokines in my experience that I've seen have visible effects on mice shortly after injection.  Expect your mice to stop most activity and huddle in a corner of the cage.  It will also give them diarrhea.  Their grooming habits will also decline (although that may be hard to notice in nude mice).  Bottom line is your probably going to make the mice feel like they have a really bad case of flu, so make sure your experiment is worth it.
My experience with dosing is with single injections.  Human TNF does not work as well on mice as mTNF.  Our maximal dose for a single injection was 2 micrograms rmTNF and 20 micrograms rhTNF.  

ltrbrown
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Carson O Genic:

Carson O Genic:
May I ask which strain(s) you use? I am learning that sensitivity to TNF-alpha is extremely strain dependent (LD100 = 12.5 to 25 ug according to Staelens et al. PMID: 12089334). This was with mTNF-alpha, of course, so I thank you for the info about h vs mTMF-alpha. I am obliged to use hTNF-alpha because we are investigating human tumor cell lines with various genetic backgrounds... perhaps that means the hTNF-alpha will be more targeted to the cancer cells.

marcus muench
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 Balb/c 

 Balb/c 
Effects of TNF on tumor growth, wow, your going back to the cachetin days.

xiaochanzm
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Hi! I am also planning to

Hi! I am also planning to work on TNF injection to mice. I want to see the septic shock of the mice and compare the mortality of wt and my transgenic mice. However, as you mentioned, this process will be very painful for the mice. I am afraid it is considered inhumane and will not be approved by the IACUC in our university. Does anyone know about anything that we can do to minimize the pain in the mice? Or are there some other reliable parameters that I can use to predict the death of the mice (like body temperature?) so that I can euthanize them in advance? Thanks very much!