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Groupe D’Etude de la Reproduction Chez le Mâle (GERM) INSERM U. 435, Université de Rennes I, Campus de Beaulieu (C.C., E.G., B.J.), 35042 Rennes, Bretagne; Rhône-Poulenc Rorer, Département de la Sécurité du Médicament, Centre de Recherche de Vitry-Alfortville (C.C., F.B.), 94403 Vitry sur Seine Cedex; INSERM Institut National de Recherche Agronomique INRA U. 418, Hôpital Debrousse (H.L., J.S.), 69322 Lyon Cedex 05, France; and Eurogenetics (E.B.), 3980 Tessenderlo, Belgium
Address all correspondence and requests for reprints to: Bernard Jégou, Groupe d’Etude de la Reproduction Chez le Mâle INSERM U 435, Université de Rennes I, Campus de Beaulieu, 35042 Rennes, Bretagne, France. E-mail: bernard.jegou{at}univrennes1.fr
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form of IL-1, whereas both
forms, and ß, are present in Leydig cells. The demonstration that human Leydig and Sertoli cells produce IL-1 and -6 under the control of gonadotropin hormones and exogenous factors, opens the possibility to study the involvement of these cytokines in the control of testis function, in normal and pathological conditions in men. 82:
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, and the neutral form, IL-1ß, and is
synthetized by a wide variety of cells (2). They are coded by two
different genes (3) but bind to the same receptor (4). Furthermore, the
rat testis contains large amounts of an IL-1-like factor (5). In rat
and mouse testicular interstitial tissue, Leydig cells and macrophages
(6, 7, 8, 9, 10) secrete IL-1, predominantly the IL-1ß form, whereas within
the seminiferous tubules, Sertoli cells and possibly germ cells produce
IL-1 (11, 12). Sertoli cell IL-1 secretion increases during sexual
maturation and fluctuates during the seminiferous epithelium cycle (11, 13). Furthermore, testicular IL-1 messenger RNA (mRNA) and protein can
be regulated by gonadotropin hormones (9, 14) and by known activators
of macrophages/monocytes IL-1, such as lipopolysaccharides (LPS) and
latex beads (9, 10, 15). One of the important functions of IL-1 is to induce the production of IL-6, another member of the cytokine family, in various cell types (16). Accordingly, within the rat testis, Sertoli cell IL-1 is known to stimulate the production of IL-6 by an autocrine mechanism, through the lipoxygenase pathway (17, 18). Sertoli IL-6 production is, like IL-1 production, stage-dependent and enhanced by LPS and FSH (17, 19). Absent in rat (17, 20) and human germ cells (our unpublished results), IL-6 mRNA and protein are, in contrast, also produced constitutively by rat Leydig cells and positively regulated by IL-1 and human CG (hCG) (20, 21).
Various actions of IL-1 and IL-6 have been observed within the testis, suggesting that these cytokines are important paracrine regulators (6, 19, 22, 23, 24, 25, 26).
In humans, the presence of IL-1 has been evidenced in testicular cytosols (27), but the cellular origin(s) and regulation of its production so far remains unknown. Therefore, the purpose of the present study was to investigate whether the two main somatic cell types of the testis, namely Sertoli and Leydig cells, are sources of IL-1 and -6 in humans, and how these cytokines were regulated. This report presents evidence of the presence of IL-1 and -6 mRNAs and immunoreactive and bioactive proteins in human adult Leydig and Sertoli cell cultures and describes their regulation by gonadotropins, LPS, and latex beads.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Testes were removed from five adult men (age, 21–45 yr) in recent cerebral death, at the same time as kidneys were removed for transplantation. This protocol was approved by the Ethical Committee of the University of Lyon, France.
Isolation and culture of human Leydig and Sertoli cells were performed according to a previously described method (28). Cells were plated in 12-well culture dishes at 106 cells/mL per well for Leydig cell preparations and 5–10 cell aggregates 106/mL per well for Sertoli cell preparations (1 aggregate gave 2.7 ± 0.7 plated cells), on extracellular basal membrane from bovine corneal endothelial cells prepared as previously described (29).
The medium was discarded and replaced after 24 h for Sertoli and Leydig cells. In some instances, a hypotonic treatment of the Sertoli cell cultures was performed with a TRIS-HCl buffer solution 20 mmol/L, pH 7.4, for 2.5 min (30) to remove most of the contaminating germ cells (see Results). The cells were further cultured for 24 h before treatment with hCG (10-9 mol/L; Organon, Paris, France) for the Leydig cells, recombinant human FSH (100 mU/mL, kindly provided by The Ares Serono group, Geneva, Switzerland) for the Sertoli cells, or LPS from E. coli (50 µg/mL; Sigma Chemical Co., St. Louis, MO) for both Leydig and Sertoli cells. In some experiments, Sertoli cells were also incubated with latex beads (7 x 108/mL; Sigma) for 24 h. At the end of the incubation, the media were collected, centrifuged, and stored at -20 C until IL-1 and -6 assays were performed, whereas cells destined for RNA preparation were collected and stored at -80 C.
Characterization of human testicular cell preparations. The
presence of Sertoli cells in the Leydig culture was checked by light
microscopic examination. Human Sertoli cells presented the typical
granulations previously described (31). Sertoli cells accounted for
less than 1% of the Leydig cell preparation. Seventy to eighty percent
of the Percoll purified cells were positive for 3ß-hydroxysteroid
dehydrogenase 
4-5 isomerase (3ß-HSD) histochemical coloration
(32).
The number of Leydig cells contaminating the Sertoli cell-enriched fraction was assessed by the 3ß-HSD histochemical coloration and found to vary between 3–6%.
The proportion of germ cells in the Sertoli cell preparations was decreased by the various washing steps of the cultures. The number of germ cells remaining in the Sertoli cell preparations averaged 8–12% at the time the cells were exposed to FSH, LPS, or latex beads. The hypotonic shock lowered this number to 1–5%. The viability of plated human Leydig and Sertoli cells, as checked by trypan blue exclusion, was > 95% at the different times of culture.
Immunocytochemical identification of macrophages. Macrophages were identified in the testicular cell preparations by immunocytochemistry with the monoclonal mouse antihuman monocyte CD14 antibody, as primary antibody in an alkaline phosphatase antialkaline phosphatase staining technique (DAKO S.A., Trappes, France). An aliquot of the Leydig and Sertoli cell-enriched preparations (105 cells in 100 µL) was cytofuged onto polylysin-coated glass slides, acetone fixed, and stored at -70 C until used. Immunocytochemical staining was performed according to manufacturer’s instructions, the anti-CD 14 was used at a dilution of 1:20. Ficoll-isolated human blood leukocytes were used as positive (stained red) control; reactions performed on testicular cells without the primary antibody were considered negative control. Hematoxylin counterstaining was performed. Macrophages (CD14+ cells) accounted for less than 0.1% of the cells in the Sertoli cell preparations and 4–8% of the cells in the Leydig cell preparations.
Isolation and culture of human blood monocyte-derived macrophages
Human blood monocyte-derived macrophages were prepared according to the method described by Audran et al. (33). Macrophage purity, analyzed with a Coulter Counter ZM (Coulter, Marseilles, France), was consistently 95% and cell viability over 95%, as judged by trypan blue staining. Macrophages were seeded at 5 x 106/mL in Iscove-modified Dulbecco’s medium and stimulated in culture for 90 min in the presence of 1 µg/mL LPS before being collected for RNA preparation and RT-PCR analysis.
Preparation of total RNA and RT-PCR analysis
Total RNA was extracted from human blood macrophages and from
human Leydig and Sertoli cell cultures, as described previously (34).
First-strand complementary DNAs (cDNAs) were synthesized from 2.5 µg
total RNA, with 10 ng/µL random hexanucleotides primers (Pharmacia
Biotech, Saclay, France) and 200 U of Moloney-murine leukemia virus
reverse transcriptase (Gibco BRL, Life Technologies, Eragny, France),
in a Tris-HCl-MgCl2 reaction buffer containing 0.5 mmol/L
of each deoxynucleotide triphosphate (Boehringher Mannheim, Meylan,
France), 10 mmol/L dithiothreitol, and 40 U of RNasin (Promega,
Charbonnières, France) in a final volume of 20 µL. The reverse
transcriptase reaction was performed for 1 h at 37 C, and the
reaction volume was then brought up to 50 µL (cDNAs RT+). Reverse
transcriptase negative controls were performed in parallel, using the
same RNA samples without Moloney-murine leukemia virus reverse
transcriptase in the reaction medium (cDNAs RT-). After RT of RNA, 3
µL of the resulting cDNA (RT+) was used for PCR, using four couples
of oligonucleotide primers (Clontech Laboratories, Ozyme,
Montigny-Le-Bretonneux, France) corresponding to positions 1038/1067
and 1876/1905 for the human ß-actin sequence (35), 205/228 and
744/768 for the human IL-1 sequence (3), 285/314 and 587/616 for the
human IL-1ß sequence (3), and 34/56 and 640/661 for the human IL-6
sequence (36). As negative controls, PCRs were carried out on cDNA
RT-. PCR was performed on a Biometra trio-thermoblock TB1 DNA Thermal
cycler (Biometra Göttingen, Germany) (94 C, 5 min followed by 35
cycles, 94 C, 1 min; 60 C, 1 min; 72 C, 2 min; and finally 72 C, 10
min). Ten microliters of the PCR products were run on a 1% agarose gel
with ethidium bromide and photographed under UV light. To verify the
specificity of the amplified PCR products visualized, we purified them
to perform specific enzymatic digestion (Geneclean II kit, Bio 101, La
Jolla, CA) as follows: the 564 bp IL-1 product was cut by
EcoRI; the 332 pb IL-1ß product was cut by
HindIII, and the 628 bp IL-6 product was cut by
Sau3AI. These three enzymes were chosen because they have
only one restriction site in the sequences amplified, as shown by the
analysis of their restriction maps (Server Bisance, CITI 2, Paris,
France).
Bioassays of IL-1 and -6
IL-1 was quantified in the culture media using the murine thymocyte proliferation assay (18, 37). The female mice C3H/HeJ used in the assay were purchased from Charles River, Wiga, Sulzfeld, Germany. One unit of IL-1 was defined as the quantity of tested material required to double [3H]thymidine incorporation into thymocytes when compared with phytohemagglutinin (PHA; DIFCO, Detroit, MI)-stimulated cultures, PHA being a comitogen factor added to all the assay. In our assay, 1 U IL-1 corresponds to 0.32 ± 0.10 ng.
IL-6 was measured using the specific IL-6-dependent 7TD1 mouse hybridoma cell line proliferation assay (17, 38). In this assay, 1 U was defined as the IL-6 concentration that gave half-maximal proliferation and corresponds to 1 ± 0.27 pg of IL-6.
The intra- and interassay coefficients of variation were <20 and <28% for the IL-1 bioassay, respectively, and <10% for both for IL-6. All samples from each independent study were run in the same IL-1 or -6 bioassay. IL-1 and -6 were undetectable in media incubated for 24 h at 35 C in extracellular basal membrane-coated culture dishes, without testicular cells.
Neutralization experiments
Because the IL-1 bioassay does not discriminate between the
IL-1ß and -1 forms, we performed neutralization experiments using
specific antibodies to identify the type of IL-1 involved in the
bioactivities measured.
Various dilutions of human recombinant IL-1 and -1ß (rhIL-1 and
-ß; Sigma) and of media from Leydig and Sertoli cultures were
incubated for 4 h at 37 C with the given concentration of: rabbit
antihuman IL-1 (0.15 mg/mL; a gift from Dr. B. Ferrua, Nice,
France); or of murine antihuman IL-1ß neutralizing antibodies (0.01
mg/mL; a gift from Dr. J. Wijdenes, Innotherapie Laboratoires,
Besançon, France); or of both anti-IL-1 and anti-IL-1ß
antibodies; or of nonimmune rabbit IgG (0.15 mg/mL); and/or of
supernatant of nonimmune hybridoma (0.01 mg/mL) used as controls. At
the end of the incubation period, all the samples were bioassayed for
IL-1.
In parallel to these experiments, we also established that the
antibodies used did not interfere with the proliferation of the
thymocytes used for the IL-1 bioassay and displayed the expected
effects on rhIL-1 (anti-IL-1 antibody neutralizing only the activity
of rhIL-1 and anti-IL1ß antibody neutralizing only the activity of
rhIL-1ß).
IL-1, -1ß, and -6 enzyme linked immunosorbent assays
(ELISAs)
Immunoreactive IL-1, -1ß, and -6 of Leydig and Sertoli cell
culture media were quantified using specific ELISA kits from
Eurogenetics (Tessenderlo, Belgium). According to the manufacturer, no
cross-reactivity was measured between: IL-1ß, -2, -3, -4, -6, tumor
necrosis factor (TNF)- or -ß, granulocyte-macrophage
colony-stimulating factor (GM-CSF) or granulocyte-colony stimulating
factor (G-CSF), and IL-1 ELISA; IL-1, -2, TNF, or interferon-
,
and IL-1ß ELISA; IL-1ß, -2, -3, -4, TNF- or -ß, GM-CSF or
G-CSF, and IL-6 ELISA. Sensitivity of IL-1, -1ß, and -6 ELISA kits
were 10, 2, and < 5 pg/mL, respectively. The intraassay
variations were <15, 7, and 10% for IL-1, -1ß, and -6,
respectively.
Because of the great difficulty in obtaining human testis and the small volumes of the culture media available for analysis, ELISAs could not be performed on all samples run in the IL-1 and -6 bioassays. Therefore, to verify the results of the bioassays, ELISA measurements were only performed on a selected number of samples of each experiment.
Statistical analyses
Results are expressed as means ± SEM. Statistical analyses were performed by the use of Student’s t test after ANOVA. Significance was defined as P < 0.05.
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Results |
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Our RT-PCR experiments demonstrated the presence of IL-1 (564
bp), IL-1ß (332 bp), and IL-6 (628 bp) mRNAs in the Leydig and
Sertoli cell preparations (Fig. 1
). The integrity of our
RNA preparations was assessed by the presence of ß-actin mRNAs in
blood-activated macrophages and in Leydig and Sertoli cell
preparations, as confirmed by the expected specific PCR product (Fig. 1). The RT-PCRs performed without reverse transcriptase were always
negative, establishing the absence of genomic contamination of our
samples (data not shown). The verification of the RT-PCR products
IL-1, -6, and -1ß by enzymatic digestion are presented in Fig. 2 and show that the 564-bp IL-1, the 332-bp IL-1ß,
and the 628-pb IL-6 product-generated fragments (Fig. 2) expected from
the restriction map analyses of the three sequences studied.
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Bioassays revealed that under basal culture conditions Sertoli
cell-conditioned media displayed both IL-1 and -6 bioactivity (Fig. 3, A and B). Exposure of the Sertoli cells to FSH
enhanced IL-6 bioactivity, but not IL-1 production. In contrast,
exposure of the cells to LPS or to latex beads greatly increased the
production of both cytokines (Fig. 3, A and B).
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Because recent studies suggested that IL-1 is also probably
produced by germ cells (rat, Ref. 12 ; humans, our unpublished
observations), and because our Sertoli cell preparations were
contaminated by these cells (8–12%), we performed a series of
experiments assaying IL-1 in media from cultures in which the number of
contaminating germ cells was reduced (1–5% remaining) by hypotonic
treatment (30). Under these basal conditions, Sertoli cells still
produced IL-1, though at levels that represented about one-third of the
levels measured in cultures without hypotonic treatment (Fig. 4A vs. 3A). Furthermore, after elimination of
germ cells, a significant stimulation of IL-1 bioactivity by FSH was
revealed, whereas the strong stimulation by LPS was even enhanced.
Unlike IL-1 bioactive levels, IL-6 bioactivity levels were only
marginally influenced by the hypotonic treatment (Fig. 4B
vs. 3B), which is consistent with our previous observations
that germ cells do not produce IL-6 (rat, Ref. 17 ; human, our
unpublished observations).
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As in Sertoli cells, bioactive IL-1 and -6 were found in
conditioned media of human Leydig cell-enriched preparations cultured
in basal conditions. Both hCG and LPS were found to stimulate markedly
the production of both these cytokines (Fig. 5, A and
B).
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The thymocyte proliferation assay used to measure IL-1 bioactivity
in this study has been shown to cross-react with several other
cytokines (39, 40) and does not discriminate between IL-1 and -1ß.
Nonimmune rabbit IgG and supernatant of nonimmune hybridoma had no
neutralizing effect on Sertoli and Leydig cell IL-1 bioactivity (data
not shown). In the absence of LPS, the IL-1 activity detected in
Sertoli cell-conditioned media was completely neutralized by
anti-IL-1 antiserum, but not affected by anti-IL-1ß antibody (Fig. 6A). As shown above (Fig. 3A
), LPS markedly stimulated
Sertoli cell IL-1 activity (Fig. 6A). Furthermore, the addition of the
anti-IL-1 antibody alone or in combination with the anti-IL-1ß
antibody reduced this activity by approximately 75%; whereas no effect
of the anti-IL-1ß antibody used alone was seen. Therefore, most
bioactivity detected in Sertoli cell media is attributable to
IL-1.
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antibody but not with
the anti-IL-1ß antibody (Fig. 6B). In LPS-stimulated Leydig cells
incubated with the anti-IL-1 antibody alone, IL-1 levels were 80%
of that measured in the controls, but the decrease observed was not
statistically significant. A much greater inhibition (40%) was seen
when the anti-IL-1ß antibody was used, clearly indicating the
presence of IL-1ß. This inhibition was even greater (70%) when both
antisera were added together (Fig. 6B). It is concluded that most
probably, in basal as well as in LPS-stimulated conditions, both
IL-1 and -1ß are present in Leydig cells. Immunological activity of IL-1 and -6 in Sertoli and Leydig cell culture media
In confirmation of the results of bioactivity presented above
(Fig. 3A), IL-1 was detected in the Sertoli cell media and was
stimulated by LPS (
3- to 5-fold) and latex beads (about 10-fold),
but not at all or only marginally by FSH (Fig. 7A). The
very low IL-1ß activity detected in these media by the ELISA kit did
not increase when the Sertoli cell preparations were incubated with
FSH, LPS, or latex beads (data not shown). Low IL-1 production was
detected in Leydig cell media (Fig. 7B). This production was enhanced
by hCG ( 2-fold) and by LPS ( 2- to 4-fold). Much higher levels
of IL-1ß were measured in these medias, which were increased in the
presence of hCG ( 2-fold) and LPS ( 3-fold) (Fig. 7C).
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and 5B): IL-6 was
constitutively produced by both Sertoli and Leydig cells and stimulated
markedly by FSH ( 5-fold), LPS ( 10-fold), and latex beads (
15-fold) in Sertoli cell-enriched preparations (Fig. 8A)
and by hCG ( 3-fold) and LPS (>2-fold) in Leydig cell-enriched
preparations (Fig. 8B).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The results of experiments using specific antibodies and immunoassays
led to the conclusion that the IL-1 produced by human Sertoli cells is
essentially the IL-1 type. Although the extremely high sensitivity
of RT-PCR allowed the detection of IL-1ß mRNA in the Sertoli cell
cultures, the IL-1ß form accounted for a very marginal part of IL-1
immunoactivity in the human Sertoli cell media. Whether the IL-1ß
mRNA originated from the very few contaminant macrophages remains to be
determined. In contrast, the immunoneutralization experiments performed
on Leydig cell media revealed that in both basal and LPS-stimulated
conditions, IL-1 bioactivity was not completely abolished by both the
anti-IL-1 and anti-IL-1ß antibodies. This probably indicated that
the Leydig cell media contained not only IL-1 and IL-1ß, the
presence of which was demonstrated by the use of the specific
antibodies taken separately and by the immunoassays, but also other
products interacting with the IL-1 bioassay. We established that IL-6
was not involved in this phenomenon, because addition of a specific
antihuman IL-6 antibody to Sertoli and Leydig cell culture media
containing the anti-IL-1 and IL-1ß antibodies did not reduce the
residual IL-1 bioactivity observed (data not shown). Moreover, a
synergistic effect of IL-2 can be excluded, because human testicular
cytosols do not exhibit IL-2 activity (27). In other respects, TNF-,
not found in media from Sertoli and Leydig cells, could originate from
contaminant macrophages and germ cells (8, 42, 43). In a similar
manner, GM-CSF, a potent costimulator factor of murine thymocyte
proliferation (40), found recently in rat testicular
macrophage-conditioned media (8), could contribute to the bioactivity
detected in Leydig cell media. Results of IL-6 immunoactivity of our
ELISA experiments were also consistent with those obtained with the
IL-6 bioassay for both human Sertoli and Leydig cell-enriched
cultures.
Our Sertoli cells were only slightly contaminated by macrophages, as assessed by the very low number of CD14+ cells detected in the Sertoli cell-enriched preparations, but they were contaminated by germ cells because of the close anatomical contact existing in vivo between the Sertoli cells and the germ cells (44). Because recent studies have shown that IL-1, but not IL-6, is present in rat and human germ cells (rat, Refs. 12, 17, 20 ; human, our unpublished observations), part of the IL-1 measured in the Sertoli cell-enriched cultures might originate from the germ cells contaminating the culture. This probably explains why production of IL-1, but not that of IL-6, was reduced after hypotonic treatment of Sertoli cells. However, it is also most probable that the decline of IL-1 could also result to some extent from the direct effect of the hypotonic treatment (45). It is well established that FSH action on the seminiferous epithelium is mediated by Sertoli cells (46). Interestingly, whereas in the presence of germ cells FSH was unable to enhance Sertoli cell IL-1 production, a significant FSH-induced stimulation of this production was observed after germ cell removal. This could be caused by the lifting of the inhibitory effect that germ cells exert on Sertoli cell responsiveness to FSH, as evidenced in different species (47).
In contrast to IL-1, IL-6 secretion was markedly stimulated by FSH, even in the presence of germ cells, as previously observed in the rat (17). The fact that, as in the rat, FSH stimulated both IL-1 and -6 production is a very strong argument to support the hypothesis that Sertoli cells are producers of both of these cytokines in the human testis. However, because of the intrinsic limitations of the techniques presently available to study human Sertoli cell IL-1 activity, it cannot be totally ruled out that a FSH-stimulated Sertoli cell factor could have stimulated a contaminating cell to produce these cytokines, though this appears very improbable, considering the low proportion of contaminants in our Sertoli cell preparations. Endotoxins from gram-negative bacteria, such as LPS from E. coli, are potent activators of cytokine production. They are able to stimulate levels of rat testicular IL-1 and -6 production in vitro (6, 10, 15, 17) and in vivo (6, 7, 48). The present study, showing an LPS-induced increase of the levels of both IL-1 and -6 in human Sertoli cell media are in line with these results. Our results also demonstrate that, as in the rat Sertoli cells (15, 17), phagocytosis of latex beads, which are other activators of macrophage IL-1 production (3), induces a strong stimulation of IL-1 and -6 release by human Sertoli cell-enriched cultures. This effect of latex beads is considered to be analogous with the residual body effect (15, 17).
It has been shown that rat Leydig cells also produce cytokines (9, 10, 20, 21). However, because the interstitium of the testis contains a high proportion of macrophages, the involvement of both Leydig cells and macrophages in cytokine production is probable. Whereas our Sertoli cell preparations were virtually uncontaminated by macrophages, Leydig cell preparations contained 4–8% of CD14+ cells. However, it is notable that hCG does not regulate macrophage function in general and macrophage IL-6 production in particular (6). Therefore, the fact that both IL-1 and -6 production are stimulated by hCG in this study, which was also observed in cultured rat Leydig cells (9, 21), very strongly suggests that human Leydig cells are producers of these cytokines. Even so, as for human Sertoli cells, it cannot totally be excluded that hCG may have stimulated a Leydig cell factor that could have induced production of IL-1 or -6 by one of the cell types contaminating our Leydig cell fraction. In addition to hCG, LPS was found to markedly stimulate IL-1 and -6 release from human Leydig cells.
In the rat, IL-1 has been found to stimulate DNA synthesis in preleptotene spermatocyte and in spermatogonia (22), whereas IL-6 was found to exert the reverse action (19), therefore suggesting major effects of these cytokines in the spermatogenetic process. Furthermore, IL-1 and -6 were also found to modulate Sertoli cell function in vitro (23, 24, 25), and IL-1 was shown to influence Leydig cell function (6), as well as to stimulate Leydig cell DNA synthesis in immature rats (26). Whether similar important IL-1 and -6 paracrine actions occur in the human testis remains to be established.
By providing the first data supporting that human Leydig and Sertoli cells produce IL-1 and -6, and that IL-1 and -6 productions are under the control of physiological (gonadotropin) and immunogenic (LPS) factors, or can be induced by the phagocytosis of latex beads by Sertoli cells, we open the way to the investigation of the role of these cytokines in the autocrine and paracrine regulations of the normal and pathophysiological testicular function in men.
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Acknowledgments |
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Footnotes |
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2 Recipient of a Rhône-Poulenc Rorer fellowship.
Received September 19, 1996.
Revised January 15, 1997.
Accepted January 23, 1997.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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messenger
ribonucleic acid expression in primary cultures of Leydig cells. Endocrinology. 129:2862–2866.[Abstract]
synthesis in rat testis. Mol Cell Endocrinol. 82:R13–R16.
is
constitutively expressed in immature male germ cells from rats. Mol
Cell Endocrinol. 105:R19–R23.
production. Biochem Biophys
Res Commun. 185:154–161.[CrossRef][Medline]
release which
triggers IL-6 production by an autocrine mechanism, through the
lipoxygenase pathway. Endocrinology. 136:3070–3078.[Abstract]
. Endocrinology. 129:1614–1620.[Abstract]
on
transferrin release from Sertoli cells in culture. Endocrinology. 129:256–262.[Abstract]
5–3ß-hydroxysteroid dehydrogenase in
testicular cell cultures. Endocrinology. 79:406–410.[Medline]
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 M. K. O'Bryan, O. Gerdprasert, D. J. Nikolic-Paterson, A. Meinhardt, J. A. Muir, L. M. Foulds, D. J. Phillips, D. M. de Kretser, and M. P. Hedger Cytokine profiles in the testes of rats treated with lipopolysaccharide reveal localized suppression of inflammatory responses Am J Physiol Regulatory Integrative Comp Physiol, June 1, 2005; 288(6): R1744 - R1755. [Abstract] [Full Text] [PDF]
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 Y Okuma, K Saito, A E O'Connor, D J Phillips, D M de Kretser, and M P Hedger Reciprocal regulation of activin A and inhibin B by interleukin-1 (IL-1) and follicle-stimulating hormone (FSH) in rat Sertoli cells in vitro J. Endocrinol., April 1, 2005; 185(1): 99 - 110. [Abstract] [Full Text] [PDF]
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 R. Le Goffic, T. Mouchel, A. Ruffault, J.-J. Patard, B. Jegou, and M. Samson Mumps Virus Decreases Testosterone Production and Gamma Interferon-Induced Protein 10 Secretion by Human Leydig Cells J. Virol., March 1, 2003; 77(5): 3297 - 3300. [Abstract] [Full Text] [PDF]
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J. Oyhenart, R. Le Goffic, M. Samson, B. Jegou, and N. Raich Phtf1 Is an Integral Membrane Protein Localized in an Endoplasmic Reticulum Domain in Maturing Male Germ Cells Biol Reprod, March 1, 2003; 68(3): 1044 - 1053. [Abstract] [Full Text] [PDF]
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L. Walch and P. L. Morris Cyclooxygenase 2 Pathway Mediates IL-1{beta} Regulation of IL-1{alpha}, -1{beta}, and IL-6 mRNA Levels in Leydig Cell Progenitors Endocrinology, September 1, 2002; 143(9): 3276 - 3283. [Abstract] [Full Text] [PDF]
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R. Le Goffic, T. Mouchel, F. Aubry, J.-J. Patard, A. Ruffault, B. Jegou, and M. Samson Production of the Chemokines Monocyte Chemotactic Protein-1, Regulated on Activation Normal T Cell Expressed and Secreted Protein, Growth-Related Oncogene, and Interferon-{gamma}-Inducible Protein-10 Is Induced by the Sendai Virus in Human and Rat Testicular Cells Endocrinology, April 1, 2002; 143(4): 1434 - 1440. [Abstract] [Full Text] [PDF]
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 J. Morales-Montor, S. Baig, R. Mitchell, K. Deway, C. Hallal-Calleros, and R. T. Damian Immunoendocrine Interactions During Chronic Cysticercosis Determine Male Mouse Feminization: Role of IL-6 J. Immunol., October 15, 2001; 167(8): 4527 - 4533. [Abstract] [Full Text] [PDF]
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S. Emiliani, M. Van den Bergh, A.-S. Vannin, J. Biramane, M. Verdoodt, and Y. Englert Increased sperm motility after in-vitro culture of testicular biopsies from obstructive azoospermic patients results in better post-thaw recovery rate Hum. Reprod., November 1, 2000; 15(11): 2371 - 2374. [Abstract] [Full Text] [PDF]
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D. Zeyse, E. Lunenfeld, M. Beck, I. Prinsloo, and M. Huleihel Induction of Interleukin-1{alpha} Production in Murine Sertoli Cells by Interleukin-1 Biol Reprod, May 1, 2000; 62(5): 1291 - 1296. [Abstract] [Full Text]
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