Tumor necrosis factor as effector molecule in monocyte mediated cytotoxicity.

نویسندگان

  • H W Ziegler-Heitbrock
  • A Möller
  • R P Linke
  • J G Haas
  • E P Rieber
  • G Riethmüller
چکیده

A newly developed assay system which uses actinomycin D (Act D) pretreated Wehi 164 target cells allows for the measurement of human monocyte cytotoxicity in a 7-h 51Cr release assay. Using the monocyte specific monoclonal antibody M42 in a direct rosetting procedure we confirm herein that among human peripheral blood mononuclear cells cytotoxicity is restricted to monocytes. When applying stringent conditions that exclude exogenous lipopolysaccharide (LPS) we could demonstrate that as little as 0.1 ng of LPS per ml triggers this cytotoxicity. Further, a factor can be detected in supernatants of mononuclear cells which is also cytotoxic against Act D treated Wehi 164 cells. This cytotoxic factor can be triggered by LPS within 4 h, but at as low a LPS concentration as 0.001 ng/ml. Since one of the LPS triggered monocyte products is tumor necrosis factor (TNF), we tested the effect of recombinant TNF cloned from the U937 cell line and we could show potent lytic activity against Act D pretreated but not, or only minimally, against untreated Wehi 164 target cells. Recombinant TNF rapidly lysed the target with significant specific release occurring as early as after 3 h in the assay. By contrast, recombinant interleukin 1 gave no lysis while lymphotoxin derived from the RPMI 1788 cell line was effective. An affinity purified antiserum directed against TNF neutralized the lytic activity of recombinant TNF and also the cytotoxic factor produced by LPS triggered mononuclear cells, while the antiserum was ineffective against lymphotoxin. Further, the antiserum when added to the assay of effector cells and Act D treated Wehi 164 cells also completely ablated cytotoxic activity. Size fractionation of cytotoxic factor and recombinant TNF by high pressure liquid chromatography led to a superimposable peak of cytotoxicity in the molecular weight range of 9,500-17,000. Further, immunoblotting with the anti-TNF antibody revealed the same Mr 15,500-16,500 band for the recombinant TNF and LPS triggered cytotoxic factor. Taken together, our data demonstrate that the cytotoxic activity of human monocytes against Act D treated Wehi 164 is mediated entirely by a LPS triggered molecule that is very similar or identical to the human tumor necrosis factor. The assay system thus provides a powerful tool to analyze the biology of TNF in humans.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Comparison of recombinant tumor necrosis factor and the monocyte-derived cytotoxic factor involved in monocyte-mediated cytotoxicity.

A direct comparison of recombinant tumor necrosis factor (rTNF) and the monocyte-derived cytotoxic factor (CF) which is involved in monocyte-mediated cytotoxicity revealed immunological, physiochemical, and biological similarities, indicating that TNF is an effector molecule in monocyte-mediated cytotoxicity. Neutralizing antiserum raised against rTNF completely inhibited the ability of CF-cont...

متن کامل

Comparison of Recombinant Tumor Necrosis Factor and the Monocyte-derived Cytotoxic Factor Involved in Monocyte-mediated Cytotoxicity1

A direct comparison of recombinant tumor necrosis factor (rTNF) and the monocyte-derived cytotoxic factor (CF) which is involved in monocyte-mediated cytotoxicity revealed immunological, physicochemical, and biological similarities, indicating that TNF is an effector molecule in monocyte-mediated cytotoxicity. Neutralizing antiserum raised against rTNF completely inhibited the ability of CF-con...

متن کامل

Monocyte-mediated Tumoricidal Activity via the Tumor Necrosis Factor–related Cytokine, TRAIL

TRAIL (tumor necrosis factor [TNF]-related apoptosis-inducing ligand) is a molecule that displays potent antitumor activity against selected targets. The results presented here demonstrate that human monocytes rapidly express TRAIL, but not Fas ligand or TNF, after activation with interferon (IFN)-gamma or -alpha and acquire the ability to kill tumor cells. Monocyte-mediated tumor cell apoptosi...

متن کامل

Human granulocyte-monocyte colony-stimulating factor and interleukin 3 stimulate monocyte cytotoxicity through a tumor necrosis factor-dependent mechanism.

Human colony-stimulating factors (CSF) exert multiple effects on the proliferation, differentiation, and function of myeloid lineage cells. In this study, the effects of three recombinant human CSFs (granulocyte-monocyte CSF [GM-CSF], interleukin 3 [IL-3], and granulocyte CSF [G-CSF]) on antibody-independent monocyte tumoricidal activity were investigated by using WEHI 164 fibrosarcoma cells as...

متن کامل

Synergistic antitumor effects of topoisomerase inhibitors and natural cell-mediated cytotoxicity.

The mechanism of augmentation of tumor cell killing by immune effector cells and chemotherapeutic drugs was studied. The effect of treating tumor cells with various antineoplastic drugs on their sensitivity to murine natural cell-mediated cytotoxicity in vitro was investigated. Pretreatment with actinomycin D at nontoxic concentrations rendered L929 and WEHI-164 tumor cells more susceptible to ...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Cancer research

دوره 46 11  شماره 

صفحات  -

تاریخ انتشار 1986