Biomedical subjects

F A Anderer

Publications and source records attributed to F A Anderer.

At least 19 recordsLinked to original sources

PubMed1996

Enhancement of human NK and LAK cytotoxicity against HCMV-infected cells by rhamnogalacturonan: specificity of reaction.

Human peripheral blood mononuclear cells (PBMC) and their subpopulations obtained from healthy donors were used to study improvement of MHC-unrestricted cytotoxic reactions against cells infected with human cytomegalovirus (HCMV) at different multiplicities of infection. Natural killer (NK) and lymphokine-activated killer (LAK) cytotoxicity against HCMV-infected cells was greatly enhanced in the presence of rhamnogalacturonan (500 ng/ml). The increase of the multiplicity of infection from MOI 0.1 to 1.0 had only a slight effect on cytotoxicity enhancement by rhamnogalacturonan. The chemical specificity of interaction of rhamnogalacturonan with effector cells and virus-infected cells was found to be analogous to the interaction with tumor cells, i.e., both types of target cells must express a receptor for rhamnogalacturonan since rhamnogalacturonan-mediated enhancement of NK and LAK cytotoxicity against HCMV-infected cells was similarly inhibited by preincubation of CD56+ effector cells with 60% deacetylated D-mannose pentaacetate.

Cells, Cultured↗

PubMed1995

Anti-(transforming growth factor beta) antibodies with predefined specificity inhibit metastasis of highly tumorigenic human xenotransplants in nu/nu mice.

Monoclonal antibodies (mAb) were prepared against conjugated transforming growth factor beta 1 (TGF beta 1) peptides: amino acid positions 48-60 and positions 86-101. Two antibodies, mAb 16-3G1 [anti-(48-60)] and mAb 5-2G6 [anti-(86-101)] cross-reacted with native TGF beta 1, -beta 2 and -beta 3 (16-3G1) or only with native TGF beta 1 (5-2G6). Both mAb were used to characterize TGF beta-mediated effects on the metastatic potential in nude mice of human carcinoma cell line SLU-1 and its metastatic subline SLU-M1. Autocrine TGF beta 1-mediated up-regulation of cell proliferation and its suppression by anti-TGF beta antibodies in vitro was recorded for SLU-M1 cells whereas SLU-1 cell proliferation in vitro appeared to be refractory to anti-TGF beta antibodies and exogenous TGF-beta 1. However, the potential of s.c. tumours to develop distant metastases in nude mice was about the same for both cell lines. Development of primary tumours and distant metastases could be suppressed by treatment of mice with anti-TGF beta antibodies. Thus we assume that the metastatic potential of tumour cells is independent of TGF beta-mediated growth-regulation effects in vitro. The anti-TGF beta-induced suppression of tumour progression and metastasis in nude mice might rather result from stimulation of the immune surveillance. TGF beta-mediated autocrine down-regulation of MHC-unrestricted cytotoxicity of activated human monocytes and CD56+ LAK cells and its reversion by anti-TGF beta antibodies could be readily demonstrated. In all our experimental series, the neutralizing potential of both anti-TGF beta antibodies, though directed against opposite sites of the TGF beta 1 molecule, was very similar.

Amino Acid Sequence↗

PubMed1995

Comparison of hexose acetate-specific receptors isolated from human leukocytes showing competitive inhibition of human CD56+NK and LAK cytotoxicity.

Specific cytotoxicity of human CD56+NK and LAK cells was quantitatively inhibited by acetylated mannose, galactose and glucose (Scand. J. Immunol., in press). The respective NK cell receptors were isolated from human leukocyte lysates by affinity chromatography based on 60% deacetylated penta-acetates of mannose, galactose and glucose. All three affinity isolates contained a main component with +/- 87 kDa molecular mass exhibiting about the same patterns of isoforms at pI 4.90, 4.75, 4.60 and 4.50 in isoelectric focusing. Moreover, preincubation of tumor target cells with the three 87-kDa receptors revealed very similar inhibitory potentials for human NK and LAK cytotoxicity showing dose-dependent inhibition between 20 (no inhibition) and 700 pmol/ml (100% inhibition) receptor concentration. The data support the assumption that the three affinity isolates contain the same type of receptor directed against a unique epitope common to acetylated mannose, galactose and glucose.

Binding, Competitive↗

PubMed1995

Optimizing production of human monoclonal IgG antibodies by in vitro-primed human PBMC: influence of CD56+ NK cell depletion.

Freshly isolated human peripheral blood mononuclear cells (PBMC) were immunomagnetically depleted of CD56+ cells. When these CD56- PBMC populations were cultured in the presence of autologous donor serum, polyclonal activation with IL-2 and pokeweed mitogen (PWM) generally resulted in exclusive production of IgG antibodies. Fusion with SP2/O-Ag14 mouse myeloma cells was highly efficient and yielded a great number of IgG-producing heterohybridomas. These conditions were used for in vitro immunization with viable human HT29 tumor cells. After fusion, an increase in hybridoma clones producing IgG monoclonal antibodies (MAb) with HT29 specificity showing a higher portion of MAb binding to the surface of viable HT29 cells was recorded. This immunizing efficiency was not observed with HT29 membrane protein fractions or HT29 proteins integrated into ISCOM particles. Investigations with human anti-alpha Gal antibodies showed that the IgG antibodies produced by the human/mouse heterohybridomas did not contain the mouse-specific Gal alpha 1-3Gal epitope.

Adjuvants, Immunologic↗

PubMed1995

Chemospecificity and cross-reactivity of target cell recognition by human CD56+ NK and LAK cells.

Inhibition of specific cytotoxicity of highly purified (> 95%) human CD56+ NK and LAK cells against K562 tumour cells was studied with various sugar acetates. Maximum inhibitory specificity was obtained with 60%-deacetylated penta-acetates of mannose, galactose, glucose, or 80%-deacetylated penta-O-acetate of N-acetyl neuraminic acid. The inhibition was strictly dosedependent and 100% inhibition was achieved in the concentration range of 500-1000 nmoles/ml with all four sugar acetate samples. Enhancement of specific cytotoxicity in the presence of rhamnogalacturonan (RG; 500 ng/ml), acting as a bridging molecule, was also inhibited in a dose-dependent manner with the same inhibitory specificity and within the same concentration range indicating involvement of the same number of sugar acetate-specific receptors. Moreover, formation of lytic CD56+ effector cell/tumour cell (E/T) conjugates was equally well inhibited whereas formation of total E/T conjugates was only partially inhibited (NK: 44-73%; LAK: 46-50%). E/T conjugate formation in the presence of RG was enhanced. Inhibition of the enhancement of formation of lytic E/T conjugates in the presence of RG was again completely accomplished with the same inhibitory specificity and within the same concentration ranges as recorded for E/T conjugate formation in the absence of RG. However, inhibition of total E/T conjugate formation was again only partially achieved at the given concentrations. The data support the assumption of an NK cell receptor with specificity for acetylated carbohydrate moieties on target cells or on bridging molecules such as RG.

Acetylation↗

PubMed1994

Enhancement of MHC-unrestricted cytotoxic activity of human CD56+ CD3- natural killer (NK) cells and CD3+ T cells by rhamnogalacturonan: target cell specificity and activity against NK-insensitive targets.

Rhamnogalacturonan-mediated enhancement of MHC-unrestricted cytotoxicity was studied with freshly isolated CD56+CD3- natural killer (NK) cells, interleukin-2 (IL-2)-activated CD56+ lymphokine-activated killer (LAK) cells und IL-2/anti-CD3-activated T cells as effector cells using NK-sensitive and NK-insensitive tumor cells as targets. The rhamnogalacturonan fractions IM, IP, and IQ were prepared from commercially available extracts of Viscum album. The dose/response relation of IM, IP, and IQ demonstrated the presence of various concentrations of cytotoxicity-enhancing compounds in all three fractions that were identified as rhamnogalacturonans by degradation studies with poly-alpha-D-galacturonidase (EC 3.2.1.15) and alpha-1,6-rhamnosidase (EC 3.2.1.40). Specific cytotoxicity of all three effector cell populations as well as the respective rhamnoagalacturonan-mediated cytotoxicity enhancement was readily inhibited in a dose-dependent manner by 60%-deacetylated mannose pentaacetate. Rhamnogalacturonan-mediated enhancement of cytotoxicity of fresh CD56+ NK cells was also observed with four of five NK-insensitive tumor cells as targets, indicating that the effector-cell/tumor-cell bridging activity of rhamnogalacturonans renders NK-insensitive targets susceptible to NK-mediated lysis. Moreover, the rhamnogalacturonan-mediated cytotoxicity enhancement became even more prominent when lymphokine-activated CD56+ LAK and CD3+ T cells were assayed with the NK-insensitive tumor cell targets.

Antigens, CD↗

PubMed1994

Identification of a mannose-acetate-specific 87-kDa receptor responsible for human NK and LAK activity.

Target cell recognition and cytotoxicity of human CD56+ NK and LAK cells is readily inhibited by acetylated mannose. Two respective NK cell receptor candidates were isolated from human leukocyte lysates by mannose acetate affinity chromatography. The 87-kDa receptor showed sequence homologies with lactoferrin and the 59-kDa receptor represented a complex of two Ca-binding proteins MRP-8 and MRP-14 reportedly expressed only by cells of myeloid origin. The 87-kDa receptor exhibited heterogeneity in isoelectric focusing and behaved entirely differently from lactoferrin. Preincubation of tumor target cells with the 87-kDa receptor inhibited competitively target cell recognition and cytotoxicity of human CD56+ NK and LAK cells.

Acetylation↗

PubMed1994

Structural specificity of MHC-unrestricted recognition of HCMV-infected target cells by human CD56+NK and LAK cells.

Structural specificity of binding and cytolysis of HCMV-infected human foreskin fibroblasts (HFF) by human NK and LAK cells was studied in inhibition assays. A sample of 60%-deacetylated alpha-D mannose penta-acetate was used as inhibitor that was previously shown to specifically inhibit binding and cytolysis of tumour target cells by human NK and LAK cells. We found now that cytolysis of HCMV-infected HFF was inhibited in a dose-dependent manner showing complete inhibition at concentrations above 640 nmoles/ml mannose acetate. This effect on cytolysis was based on inhibition of conjugate formation between virus-infected cells and CD56+NK and LAK cells. In the presence of mannose acetate (640 nmoles/ml) conjugate formation of virus-infected cells was suppressed down to the level of uninfected cells. The latter showed residual conjugate formation on the basis of adhesive interactions with chemospecifity other than for mannose acetate, which were not capable of triggering cytolytic reactions. Coculturing of target cells with LAK cells appeared to induce expression of additional mannose acetate-specific target sites yielding increases of conjugate formation and cytolysis.

Antigens, CD↗

PubMed1993

Anti-epidermal growth factor receptor monoclonal antibodies affecting signal transduction.

Monoclonal antibodies prepared against tyrosine phosphorylated epidermal growth factor receptor (EGFR) were tested for their effects on transmembrane signal transduction in A431 tumor cells. Monoclonal antibodies (mab) defined by SDS-sensitive epitopes, i.e., epitopes with conformational specificity, were most effective. Mab 5-125 reacting with a site of the extracellular EGFR domain blocked EGF-binding and cell proliferation in vitro, as well as tumor growth in vivo. However, this mab appeared not to be internalized upon binding to EGFR and did not trigger EGFR autophosphorylation. In contrast, mab 5-D43, also defined by an SDS-sensitive epitope and reacting with an extracellular EGFR site, did not block EGF binding but was readily internalized after binding to EGFR of untreated A431 cells. This mab induced EGFR tyrosine phosphorylation in cell lysates and tyrosine-specific autophosphorylation of insolubilized EGFR immune complexes. Cell growth in vitro was greatly stimulated in the presence of mab 5-D43. Since interaction of mab 5-D43 with EGFR induced most EGF-specific functions, although it did not bind to the EGF-specific site of EGFR, we have to assume that binding of mab 5-D43 to EGFR induced a conformational shift that activated the cytoplasmic EGFR kinase site. On the other hand, activation and/or accessibility of the EGFR kinase site could be blocked by mab 1-594, which is defined by an SDS-insensitive protein epitope of the cytoplasmic EGFR domain. Blocking of the EGFR kinase site by mab 1-594 also abolished EGF-induced tyrosine phosphorylation of endogenous cellular substrates with molecular masses of 145, 97, 85, 37, and 32 kDa, as well as of exogenous substrates such as GAT copolymer.

Animals↗

PubMed1993

Mechanism of stimulation of human natural killer cytotoxicity by arabinogalactan from Larix occidentalis.

Cultures of human peripheral blood mononuclear cells (PBMC) as well as cultures of preseparated peripheral non-adherent cells (PNAC) and monocytes showed enhancement of natural killer (NK) cytotoxicity against K562 tumor cells when pretreated with arabinogalactan from Larix occidentalis for 48-72 h. Lack of enhanced responses of PBMC (37% of donors) did not necessarily mean that PNAC and monocyte cultures were also non-responsive to arabinogalactan treatment. Moreover, PBMC, PNAC and monocytes of individual donors could exhibit various responses to arabinogalactan when cultures derived from bleedings after intervals of several months were assayed. Arabinogalactan-mediated enhancement of NK cytotoxicity was not initiated directly but was found to be governed by the cytokine network. Generally, arabinogalactan pretreatment induced an increased release of interferon gamma (IFN gamma), tumor necrosis factor alpha, interleukin-1 beta (IL-1 beta) and IL-6 but only IFN gamma was involved in enhancement of NK cytotoxicity since cytotoxicity enhancement of PBMC and PNAC but not that of monocytes could be blocked when anti-IFN gamma antibodies were present during pretreatment. The presence of anti-IL-2 antibodies completely blocked NK cytotoxicity enhancement of PBMC and only moderately that of PNAC and monocytes. This blocking effect was also observed when no detectable increase of IL-2 release could be recorded. The receptor specificity of arabinogalactan is not well characterized. Initial information obtained from comparative studies indicated that arabinogalactan presumably interacts with a receptor that showed specificity for a NK-cytotoxicity-enhancing oligo-saccharide from Viscum album extracts since the action of both components was not synergistic but rather competitive.

Cytokines↗

PubMed1993

Activation of human monocyte/macrophage cytotoxicity by IL-2/IFN gamma is linked to increased expression of an antitumor receptor with specificity for acetylated mannose.

Spontaneous cytotoxicity of human monocytes (purity: 92-95%) against K562 tumor cells was only observed in 31% healthy donors but, in the presence of rhamnogalacturonan (500 ng/ml), enhanced cytotoxicity was recorded for 79% (n = 14) of the donors. Monocytes activated by culturing with interleukin-2 and/or IFN gamma showed increased antitumor cytotoxicity against K562 tumor cells in 86% (n = 21) of the donors exhibiting additional increases in specific cytotoxicity when the cytotoxicity assays were carried out in the presence of rhamnogalacturonan. Increases of monocyte cytotoxicity achieved by activation with cytokines coincided with increased formation of monocyte/tumor cell conjugates. Similarly, increased monocyte cytotoxicity mediated by rhamnogalacturonan also correlated with increased monocyte/tumor cell conjugate formation most likely due to effector cell/target cell bridging as was originally described for rhamnogalacturonan interacting with CD56+ natural killer or lymphokine-activated killer cells and tumor cells. The chemospecificity of the monocyte-based receptors responsible for cytotoxicity and for monocyte/tumor cell conjugate formation, as well as for their rhamnogalacturonan-mediated enhancements, appears to be identical since all these effects could be inhibited in a dose-dependent manner by partially deacetylated (60%) mannose pentaacetate.

Cytotoxicity, Immunologic↗

PubMed1992

The CySF-L2 factor from dialysable human leucocyte extract activates natural killer cytotoxicity by induction of interferon gamma.

The mechanism of natural killer (NK) cytotoxicity activation of human peripheral blood mononuclear cells (PBMC) by CySF-L2 was elucidated. CySF-L2 is a cytotoxicity-stimulating factor isolated from dialysable human leucocyte extract, which activates NK cytotoxicity against NK-sensitive and insensitive tumour cells (K562; Daudi; Raji; MOLT4) when preincubated with effector cells for 72 h. CySF-L2-mediated activation was synergistic to interleukin-2(IL-2)-mediated activation of NK cytotoxicity. Induction of interferon gamma (IFN gamma) release was the crucial step during CySF-L2-mediated NK cytotoxicity activation since enhancement of NK activity was completely blocked when anti-IFN gamma antibodies were present during treatment of PBMC. Anti-IFN alpha, anti-TNF alpha (tumour necrosis factor alpha) anti-IL-1 and anti-IL-2 antibodies showed no blocking effect. Analysis of the supernatant culture medium after 72 h incubation of PBMC and their highly purified subpopulations demonstrated that CySF-L2 induced release of IFN gamma from CD3+T cells and CD56+CD3- NK cells and of TNF alpha and prostaglandin E2 from monocytes. CySF-L2 was also capable of activating NK cytotoxicity of highly purified (98%) CD56+CD3- NK cells as well as of monocytes (94% pure). Cell cooperation studies connected with analysis of cytokine release and enhancement of NK cytotoxicity indicated that CySF-L2 might play an essential role in the up and down regulation of NK cytotoxicity by the cytokine network.

Antigens, Surface↗

PubMed1992

The ontogeny of bovine thymostimulin production in fetal and postnatal age.

The development of thymostimulin production in the bovine fetal thymus was determined, starting at month 2 of gestation until birth. Production of fetal thymostimulin, identified electrophoretically as a peptide with 4-5000 Da, started at month 4 of gestation and achieved its maximum expression three months after birth, followed by a rapid decrease until month 18. Thymus of fetuses from the early gestational phase (2-3 months) yielded no electrophoretically detectable thymostimulin band. Biological activity of the fractions, determined by increased E-rosetting, fairly correspond to the content of the 4-5000 Da peptide moiety.

Age Factors↗

PubMed1990

Synergistic action of a plant rhamnogalacturonan enhancing antitumor cytotoxicity of human natural killer and lymphokine-activated killer cells: chemical specificity of target cell recognition.

The spontaneous natural killer (NK) and lymphokine-activated killer (LAK) cytotoxicity of highly purified CD56+CD3- NK cells (90 to 95%) against NK-sensitive and NK-insensitive target cells was drastically enhanced when a rhamnogalacturonan contained in a commercially available Viscum album extract was present during 4-h cytotoxicity assays. This enhancement correlated strictly with an increased formation of NK cell or LAK cell/tumor cell conjugate formation. Information on the chemical specificity of NK cell and LAK cell interaction with target cells and with the rhamnogalacturonan was obtained from inhibition studies. The most efficient inhibitors (100% inhibition at 5 mg/ml) were acetylated D-mannose and acetylated L-mannonic acid gamma-lactone. They specifically inhibited in a dose-dependent manner: (a) the cytotoxicity of NK cells against K562 cells and the formation of NK cell/K562 cell conjugates; (b) the cytotoxicity of LAK cells against K562 cells and Daudi cells as well as the formation of LAK cell/K562 cell and of LAK cell/Daudi cell conjugates; and (c) the synergistic effects of the rhamnogalacturonan in the cytotoxicity assays and the target cell-conjugate formation assays with NK cells and LAK cells. The inhibitory effects observed after pretreatment of NK cells or LAK cells with acetylated mannose were completely reversible, but that obtained with acetylated mannonic acid gamma-lactone was only partly reversible, and the degree of reversibility depended on the inhibitor concentration applied during pretreatment. Nonacetylated mannose or mannose derivatives up to concentrations of 20 mmol showed no inhibitory effects. A mechanistic model representing the interaction of NK cells and LAK cells with target cells and with rhamnogalacturonan is proposed.

Antigens, Differentiation, T-Lymphocyte↗

PubMed1990

Preservation of cytotoxic function during multi-cycle immunomagnetic cell separations of human NK cells using a new type of magnetic bead.

The isolation from human peripheral blood lymphocytes of natural killer (NK) cell populations by a novel magnetic cell sorting (MACS) procedure yielded large amounts of viable cells with active cytotoxic function. Non-adherent B cell-depleted lymphocytes were sequentially labelled with specific monoclonal antibodies, biotin-conjugated second antibody, FITC-conjugated streptavidin and biotin-conjugated magnetic particles (diameter 50-150 nm). In the magnetic field of a permanent magnet, positively labelled cells were retained on columns with a ferromagnetic matrix. When OKT3 was used for the depletion, 96-99% of the T cells were removed. The resulting non-labelled NK cell population contained 78-89% Leu11b+ and 87-96% Leu19+ cells. Magnetic retention of NK cells mediated by anti-Leu19 yielded about 81% and retention mediated by anti-Leu11b about 80% of total cells as determined by positive fluorescence. The resulting labelled and unlabelled cell subpopulations maintained their full NK activity as determined in 4 h cytotoxicity assays against human K562 tumor cells. The viability of non-labelled cells was fully preserved, whereas that of labelled cells slowly decreased with increasing numbers of preparative cycles. Furthermore, the ability of the isolated NK cells to show enhancement of their NK cytotoxicity after preincubation with IL-2 was maintained. The cytotoxic function of NK cells was also preserved when two or more MACS cycles using the same or different antibodies were carried out. The saving of time and the physiological condition of the isolated cells offer valuable advantages over FACS procedures.

Antibodies, Monoclonal↗

PubMed1990

A Viscum album oligosaccharide activating human natural cytotoxicity is an interferon gamma inducer.

Commercial Viscum album extract Helixor-M contains a dialysable oligosaccharide (HM-BP) that activates natural killer (NK) cytotoxicity against K562 tumour cells when preincubated with human peripheral blood mononuclear cells (PBMC) for 72 h. The activated effector cells were exclusively found in the monocyte/macrophage subpopulation. However, when peripheral non-adherent cells (PNAC) were preincubated with HM-BP for 72 h the NK cytotoxicity of CD56+CD3- NK cells was activated. This discrepancy was found to be due to the release of prostaglandin E2 from activated monocytes/macrophages, which blocked activation of the cytotoxicity of NK cells. Analysis of the supernatant culture medium after 72 h preincubation demonstrated that HM-BP induced release of interferon gamma (IFN gamma) from T cells (preferentially from CD3+CD4+ cells) and of tumour necrosis factor alpha (TNF alpha) from monocytes/macrophages. Release of IFN gamma was the crucial step for activation of NK cytotoxicity since enhancement of NK cytotoxicity during pretreatment of PBMC or PNAC with HM-BP was completely blocked in the presence of anti-IFN gamma antibodies. Anti-interleukin-2, anti-TNF alpha or anti-IFN alpha antibodies had no effect on the HM-BP-induced enhancement of NK cytotoxicity. The activation of the NK cytotoxicity of nonadherent cells by interleukin-2 treatment was found to be synergistic to the enhancement of NK cytotoxicity by treatment with HM-BP.

Cells, Cultured↗

PubMed1989

Interleukin 2-induced protein phosphorylation/dephosphorylation in human non-adherent peripheral blood mononuclear cells: initial phosphorylation of two 75-kDa proteins.

The kinetics of protein phosphorylation/dephosphorylation induced by IL-2 in cells of the non-adherent subpopulation of human peripheral blood mononuclear cells after pretreatment with and without phytohaemagglutinin (PHA) was analyzed by two-dimensional gel electrophoresis. In cells not pretreated with PHA, IL-2 induced the phosphorylation of two 75-kDa proteins with pI values 6.6 and 6.9, detectable 30 min after addition of IL-2, and the dephosphorylation of a 94-kDa (pI 4.0) protein, two 85-kDa (pI 5.2 and 5.4) proteins and a 65-kDa (pI 4.9) protein. The latter three phosphoproteins were found to be unlabelled after PHA pretreatment, but upon IL-2 stimulation the 94-kDa and the 85-kDa proteins became labelled simultaneously with two minor phosphoproteins of 68 kDa (pI 5.7) and 37 kDa (pI 4.8). Moreover, PHA pretreatment of cells induced a drastic phosphorylation of a 48-kDa (pI 6.5) and a 46-kDa (pI 6.7) protein, which were gradually dephosphorylated after IL-2 addition. Phosphorylation of the 75-kDa proteins could not be detected when the cells were pretreated with PHA prior to labelling. These results suggest that IL-2-induced phosphorylation of the 75-kDa proteins is one of the early events in IL-2 stimulation, an event already completed in PHA-pretreated cells since PHA is known to induce release of IL-2. Furthermore, the retarded appearance of the labelled 75-kDa proteins suggests an IL-2-induced de novo synthesis, possibly reflecting the expression of the 75-kDa alpha chain of the IL-2 receptor.

Antibodies, Monoclonal↗

PubMed1989

A dialysable acid factor from human leukocyte extracts activates tumor cell lysis mediated by human monocytes and natural killer cells.

Dialysable human leukocyte extract contains an acid natural killer (NK) cytotoxicity-stimulating factor, CySF-L1, which can be enriched by ion exchange chromatography. NK-cytotoxicity of human peripheral blood mononuclear cells (PBMC) against human K562 and HT29 tumor cells was strongly enhanced after 72 h pre-incubation with the factor. The CySF-L1-specific stimulation of PBMC required the presence of monocytes. The cytotoxic effector cells activated during preincubation of PBMC with CySF-1 were identified as monocytes and as NK cells present in the fraction of large granular lymphocytes (LGL). Selective cell depletion studies with LGL containing subpopulations (free of monocytes) indicated that the activated NK cells express CD 16 (Leu 11) and CD8 (T8) markers and the majority of them also the Leu7 marker. Analysis of the changes of surface marker expression of human PBMC during preincubation with CySF-L1 revealed an efficient stimulation of CD8 and TfR (transferrin receptor) expression, partly in conjunction with diminished expression of CD4 (T4). In vivo application of CySF-L1 after tumor challenge induced reduction of the incidence of tumor take and tumor development in mice.

Adult↗