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PubMed1997

Role of protein kinase A in collagenase-1 gene regulation by prostaglandin E1: studies in a rabbit synoviocyte cell line, HIG-82.

Gene expression of the matrix-degrading enzyme collagenase-1 in rabbit synoviocytes and human fibroblasts is down-regulated by prostaglandin E1 (PGE1) through a cyclic adenosine monophosphate (cAMP)-dependent pathway. In the current study, we examined the role of protein kinase A (PKA) in the PGE1-mediated effect on collagenase-1 gene expression. Collagenase-1 gene expression was rapidly induced several-fold above control both by a phorbol ester, 12-o-tetradecanoyl phorbol 13 acetate, and interleukin-1 beta (IL-1 beta) in HIG-82 synoviocytes. Treatment with PGE1 and forskolin increased PKA activity in the HIG-82 cells within 15 minutes of adding the stimulating agents. Two inhibitors of PKA, the isoquinoline-sulfonamide derivative, H-89 and a cAMP analog, RpcAMP, blocked the ability of PGE1 to down-regulate collagenase-1 gene expression. However, if PGE1 was added from 6 h to 30 minutes before the PKA inhibitor H-89, collagenase-1 gene expression was inhibited. Constitutive PKA activity was increased in HIG-82 synoviocytes stably transfected with an expression vector pCMV.C alpha that caused the HIG-82 cells to overexpress an active catalytic subunit of PKA. Cells stably transfected with an inactive, mutated C-alpha-variant showed no change in PKA activity. Collagenase-1 mRNA levels in TPA-stimulated cells were reduced to baseline levels in the pCMV.C alpha but not in the mutated C-alpha-transfected cells. These data show the importance of PKA in regulating collagenase-1 gene expression in a synoviocyte cell line.

Alprostadil↗
PubMed1998

Mechanism of mechanically induced intercellular calcium waves in rabbit articular chondrocytes and in HIG-82 synovial cells.

Intercellular communication through gap junctions allows tissue coordination of cell metabolism and sensitivity to extracellular stimuli. Intercellular Ca2+ signaling was investigated with digital fluorescence video imaging in primary cultures of articular chondrocytes and in HIG-82 synovial cells. In both cell types, mechanical stimulation of a single cell induced a wave of increased Ca2+ that was communicated to surrounding cells. Intercellular Ca2+ spreading was inhibited by 18alpha-glycyrrhetinic acid, demonstrating the involvement of gap junctions in signal propagation. In the absence of extracellular Ca2+, mechanical stimulation induced communicated Ca2+ waves similar to controls; however, the number of HIG-82 cells recruited decreased significantly. Mechanical stress induced Ca2+ influx both in the stimulated chondrocyte and HIG-82 cell, but not in the adjacent cells, as assessed by the Mn2+ quenching technique. Treatment of cells with thapsigargin and with the phospholipase C (PLC) inhibitor U73122 blocked mechanically induced signal propagation. These results provide evidence that in chondrocytes and in HIG-82 synovial cells, mechanical stimulation activates PLC, thus leading to an increase of intracellular inositol 1,4,5-trisphosphate. The second messenger, by permeating gap junctions, stimulates intracellular Ca2+ release in neighboring cells. It is concluded that intercellular Ca2+ waves may provide a mechanism to coordinate tissue responses in joint physiology.

Animals↗
PubMed2000

Comparison of HIG scintigraphy and bloodpool scintigraphy using HDP in arthritic joint disease.

AIM: Because of a similar tracer accumulation, we assumed to get the same information about synovitis in arthritic joint disease with HIG scintigraphy and bloodpool scintigraphy using HDP. Therefore, we compared retrospectively 23 patients. METHODS: In HIG scintigraphy, synovitis was diagnosed according to increasing activity from early to late image. In bloodpool scintigraphy according to an increased activity in comparison to the surrounding tissues. RESULTS: In 694 joints comparison of both scintigraphic modalities was possible, resulting in a 2 x 2 kappa coefficient of 0.93 or 0.97 by using late-phase bone scintigraphy as an anatomical marker. For intra- and interobserver agreement, 2 x 2 kappa coefficients of 0.93 and 0.88 in HIG scintigraphy, respectively 0.96 and 0.90 in blood-pool scintigraphy were calculated. CONCLUSION: This study shows an excellent agreement in the visualization of synovitis by HIG and bloodpool scintigraphy. Because of its higher objectivity and lower cost, investigation of synovitis should be performed by bloodpool scintigraphy.

Adult↗
PubMedDate not supplied

Is 99mTc human immunoglobulin G scintigraphy (HIG-scan) useful for the detection of spinal inflammation in ankylosing spondylitis?

OBJECTIVE: To determine the effectiveness of technetium-99m labelled polyclonal human immunoglobulin G (HIG-scan) scintigraphy to detect spinal inflammation in patients with ankylosing spondylitis. METHODS: Six ankylosing spondylitis (AS) patients with inflammatory axial pain and 3 spinal osteoarthritis (OA) patients with mechanical axial pain underwent an HMDP scan and an HIG-scan. RESULTS: In both AS and spinal OA patients complaining of axial back pain, mechanical and inflammatory axial pain, bone scintigraphy revealed foci of markedly increased tracer accumulation. An HIG-scan of the region of increased uptake on bone scintigraphy did not reveal increased tracer accumulation in either group of patients. CONCLUSION: The data suggest that the HIG-scan is not useful in detecting inflammatory spinal lesions in ankylosing spondylitis.

Adult↗
PubMed1997

[Imaging of early carotid artery atherosclerotic lesions with 111In-labeled polyelonalhuman IgG (HIG)].

AIM: To assess the value of scintigraphy with 111In-HIG for diagnosis and evaluation of the stage and the clinical extent of carotid artery disease in humans a prospective clinical comparative trial of scintigraphy vs. sonography was performed. METHODS: 58 patients (38 male, 20 female; mean age 60 +/- 7 years) with hyperlipidemia and ultrasonographically detectable carotid artery lesions were studied. After i.v. injection of 18.5 MBq 111In-HIG, anterior scintigraphic images of the neck were acquired. Real time two-dimensional B-mode ultrasonography of the left and the right carotid arteries was performed. RESULTS: 111In-HIG-scintigraphy as compared to the morphological gold standard (ultrasonography) had a sensitivity of 70-73%, specificity of 33-41% and a positive predictive value of 77-82% for detecting carotid atherosclerotic lesions. There was, however, no significant correlation between scintigraphy and ultrasonography. CONCLUSION: However, the data provide evidence that the two imaging techniques are visualizing different aspects of atherogenesis. On the one hand a functional one reflecting the activity of the disease (111In-HIG) and on the other hand the morphological one resembling the extent of the disease (ultrasonography).

Aged↗
PubMed2001

Specific protein-DNA and protein-protein interaction in the hig gene system, a plasmid-borne proteic killer gene system of plasmid Rts1.

The hig (host inhibition of growth) gene system of plasmid Rts1 belongs to the plasmid-encoded proteic killer gene family. Among the proteic killer genes described so far, hig is unique in that the toxin gene (higB) exists upstream of the antidote gene (higA). There are two promoters in the hig locus, Phig and PhigA, and only the former, which expresses both higB and higA genes, is negatively controlled by HigA and HigB proteins. In this study, we purified HigA protein by means of GST fusion. The electrophoretic mobility shift assay using the purified protein revealed that HigA specifically bound to the Phig region, but not to PhigA. The HigA-binding sequence was determined by DNase I footprinting assay to be a 56-bp sequence that completely covered the -35 and -10 boxes of Phig. The presence of two inverted repeats in the binding sequence and the identification of a dimer form of HigA by cross-linking experiment suggested that the protein bound to the Phig region as a dimer. HigB was purified as a GST fusion protein as well, though it was achieved only in the presence of HigA. HigA and GST-HigB formed a highly stable complex where the two proteins were present in an equimolar ratio.

Amino Acid Sequence↗
PubMed1983

Chemical linkage of erythrocytes and viral antigen in the hemolysis-in-gel (HIG) test for viral antibodies.

The sensitivity of the hemolysis-in-gel (HIG) test with rubella antigen is not improved by chemical linkage of the virus to the erythrocytes, and after such modification, IgM specific antibodies are not detectable. In the influenza HIG test with tetraazotized o-dianisidine (TOD), chromic chloride and potassium periodate as coupling reagents, increased sensitivity was observed with allantoic fluid of infected eggs as antigen. If Tween-ether treated hemagglutinin is used in the HIG test, zones of hemolysis are detectable only after treatment of the erythrocytes with TOD, chromic chloride and potassium periodate.

Allantois↗
PubMed2002

Tc-99m-labeled human polyclonal immunoglobulin G (HIG) scintigraphy in Sjögren's syndrome.

OBJECTIVE: To evaluate the usefulness of Tc-99m-HIG scintigraphy in patients with Sjögren's syndrome. METHODS: Twelve consecutive patients with verified secondary Sjögren's syndrome were included in this prospective study. The control group consisted of seven patients with Lupus erythematosus; none of them showed clinical signs of Sjögren's syndrome. Planar and SPECT images of the head were performed six hours after i.v. administration of Tc-99m HIG. RESULTS: Eleven out of twelve patients with secondary Sjögren's syndrome showed a positive result, while one was false negative. Tracer accumulation in patients with positive scintigraphy varied. All patients of the control group were negative. CONCLUSION: Our data in a limited number of patients suggest that Tc-99m HIG scintigraphy could be a modality with high sensitivity and specificity for the diagnosis of Sjögren's syndrome and can provide objective information on the severity of the disease.

Adult↗
PubMed1996

Scintigraphic evaluation of the severity of inflammation of the joints with 99TCm-HIG in rheumatoid arthritis.

A prospective study was carried out to determine the usefulness of 99TCm-human immunoglobulin G (HIG) scintigraphy in the assessment of the severity of joint inflammation. Twenty-four patients with rheumatoid arthritis were studied. The presence or absence of pain and/or swelling was evaluated in 34 joints and a clinical index taking into account the surface area of each joint was calculated. We measured the following biological markers of inflammation activity: erythrocyte sedimentation rate, C-reactive protein, haemoglobin, platelet count, serum levels of IL-6, TNF-alpha and soluble receptors of IL-2. Scintigraphic was performed 4 h after the injection of 740 MBq 99Tcm-HIG. The scans were evaluated by visual and quantitative analysis and the scores in each joint were weighted for joint size. Pathological uptake of the radiopharmaceutical was noted in 46% (24/52) of joints evaluated as painful, 89% (146/164) of swollen joints and 94% (78/83) of both painful and swollen joints. Both the visual and the quantitative scintigraphic indices correlated significantly with the clinical index, the number of painful joints, the number of swollen joints and several biological markers of inflammation. A very high correlation was also found between the visual and the quantitative scintigraphic indices (r = 0.91, P < 0.0001). In conclusion, 99Tcm-HIG scintigraphy is an objective test to detect synovitis and to assess the severity of inflammation. A careful visual analysis of scans is good enough for routine evaluations and computer quantitative analysis should be used when more accurate intra-individual variation is required.

Adult↗
PubMed1980

Direct HIG test for rubella antibodies.

A modification of the haemolysis-in-gel test for rubella is described. This involves using, instead of sera, whole blood samples taken from the earlobe and placed directly onto the test plates without any pretreatment. A comparison was made between this method and the HI test. Samples from 461 individuals were tested by both methods. The results of these two tests were in good agreement. No false negatives or false positives were observed by the direct HIG test. The direct HIG test is particularly well suited for screening large groups for rubella antibodies in connection with a vaccination program.

Adult↗
PubMed1998

Gene transfer of type 1 interleukin-1 receptor extracellular-domain complementary DNA into rabbit synovial cell line HIG-82 results in cellular blockade of interleukin-1 signal transduction.

OBJECTIVE: To produce, by means of expression cloning, a soluble type 1 interleukin-1 receptor (sIL-1R), and to assess its inhibitory properties on the IL-1 pathway. METHODS: High-affinity IL-1R sites were identified in a human chondrosarcoma cell line by means of 125I-IL-1beta binding. A 1-kilobase complementary DNA (cDNA) encoding the ligand-binding domain of the type 1 IL-1R was cloned by using polymerase chain reaction, and the cDNA was inserted into a mammalian expression vector pRc/CMV. The sIL-1R expression vector was transfected into a rabbit synovial cell line (HIG-82) and a stably transfected cell population was selected. The production of sIL-1R was confirmed in the medium of transfected cells using 125I-IL-1beta binding. 35S labeling of transfected cultures, followed by immunoprecipitation and gel electrophoresis, was used to characterize the size of the recombinant sIL-1R. Stromelysin and IL-1alpha steady-state messenger RNA (mRNA) levels were assessed by Northern blotting. Prostaglandin E2 (PGE2) release was measured by enzyme-linked immunosorbent assay. RESULTS: IL-1R on the surface of HIG-82 cells bound 125I-IL-1beta with an equilibrium dissociation constant (Kd) of 67.3 +/- 7.8 pM (mean +/- SD). Transfection of the sIL-1R expression vector into a synovial cell line in vitro resulted in the appearance of an sIL-1R protein that bound 125I-IL-1beta with high affinity in the medium (Kd = 108 +/- 5 pM). Two protein bands (Mr 42 kd and 47 kd) were immunoprecipitated with an antibody against type 1 T cell-derived sIL-1R. Expression of sIL-1R was accompanied by a marked decrease in both stromelysin and IL-1alpha steady-state mRNA levels. In conjunction, there was a significant inhibition of basal and IL-1-stimulated PGE2 released by sIL-1R-producing cells. CONCLUSION: The data suggest that gene transfer of type 1 sIL-1R into the synovium may be an effective means of inhibiting IL-1-induced metalloproteinase expression and inflammatory responses.

Animals↗
PubMed1996

A new plasmid-encoded proteic killer gene system: cloning, sequencing, and analyzing hig locus of plasmid Rts1.

A new proteic killer gene system, hig, was identified on the plasmid Rts1. The hig locus consisting of a higA and higB is directly related to the temperature sensitive host cell growth conferred by Rts1. We proved that higB encoding presumably a 92-amino-acid polypeptide inhibited segregation of plasmid free cells, and higA encoding a 104-amino-acid polypeptide suppressed the higB function both in cis and in trans.

Amino Acid Sequence↗
PubMed1996

Comparison between scintigraphy with polyclonal immunoglobulin (99Tcm-HIG) and physical examination in polyarthritic disease.

99Tcm-labelled polyclonal human immunoglobulin 99Tcm-HIG scintigraphy has been suggested as a technique to detect joint inflammation in arthritic disorders. Scintigraphy was performed in fifteen patients with active polyarthritis. All joints except the hips were scored clinically for swelling and pain by a rheumatologist and scintigraphic images were obtained at 30 minutes and four hours after injection of 350MBq 99Tcm-HIG. The images were assessed by a nuclear medicine physician according to a four grade scale. The images were easy to assess. There was a highly significant correlation between swelling and scan score, but no correlation between pain and scan score. The mechanism for the accumulation of activity to inflamed synovial tissue remains unclear. The mean values of the scan score however increased significantly between 30 minutes and 4 hours, indicating an active binding mechanism. The method has a potential as an objective tool in monitoring rheumatic diseases.

Adult↗
PubMed1991

Biokinetics and estimation of dose from 99mTc-labelled polyclonal human immunoglobulin (HIG).

Recently, polyclonal unspecific human immunoglobulin was suggested as a new agent for the localization of inflammatory lesions. Little information about the biodistribution of this radiopharmaceutical was reported so far. To obtain further information, 99mTc-labelled human immunoglobulin (HIG) was administered to a volunteer with presumed normal biokinetics. The absorbed doses to the organs were calculated according to the MIRD concept. The scintigraphic images at 1, 2, 4 and 24 h post injection demonstrated a prolonged activity retention in the blood and high activity in the kidneys, bladder and also in the liver. No significant uptake in the bowels and the marrow could be observed over the whole period of study. 27.4% of the administered activity was excreted in the urine within 24 h. The calculated organ absorbed doses, all in microGy/MBq, were estimated as follows: whole body 2.7, liver 7.3, spleen 12.0, kidneys 15.3, lungs 3.2, marrow 9.6 and gonads 17.0. From these results an effective dose equivalent of 7.9 x 10(-3) mSv/MBq was calculated. The cancer risk estimate of 5 x 10(-5), using 370 MBq 99mTc-HIG, may be considered quite low in comparison to other scintigraphic methods of diagnosing inflammation.

Abdomen↗
PubMed1998

Evaluation of 99Tcm-HIG as a lymphoscintigraphic agent in rabbits.

In this pilot study, 99Tcm-labelled human immunoglobulin G (99Tcm-HIG) was evaluated as a lymphoscintigraphic agent in five rabbits. It was injected intradermally into the web space of the hind legs of the rabbits (37 MBq/0.1 ml). Sequential scintigrams were obtained using a gamma camera for 120 min. The injection site and the hind legs were massaged post-injection. Blood samples were obtained at 5, 15, 30, 60, 90 and 120 min. Two of the rabbits were killed after 2 h. Their organs were weighed and tissue specimens were obtained, weighed and counted against a standard using a gamma counter. The lymph channels and the lymph nodes were well visualized on the scintigrams. The background activity was very low, making interpretation easier. About 30% of the injected dose migrated from the injection site by 2 h. The mean popliteal lymph node uptake was 5.71 +/- 4.62% per gram of tissue. The lymph node to other tissue concentration ratios were very high, ranging from 63:1 for the kidneys to 1099:1 for the heart. We conclude that 99Tcm-HIG is a promising new agent for the visualization of the lymphatic system due to its easy labelling procedure, the stability of the label, its widespread availability and good image quality. It may potentially be useful in detecting and evaluating inflammatory lymph nodes.

Animals↗
PubMed2006

Osteoid osteoma simulating osteomyelitis: differentiation with Tc-99m HIG scintigraphy.

We present a case of osteoid osteoma with a history of mild pain, local swelling and point tenderness on the right lower leg. The diagnosis of osteoid osteoma was difficult due to the atypical clinical history and misleading radiological and bone scan findings. When it is difficult to differentiate an osteoid osteoma from osteomyelitis using CT, MRI or bone scan; HIG scintigraphy can be used to exclude an infection.

Adolescent↗
PubMed1981

[Reasons for repeated serologically unsuccessful (HAI-HIG test) rubella vaccination (author's transl)].

Following vaccination against rubella, persons are sometimes detected who, despite repeated rubella vaccinations (Cendehill strain), show absence of seroconversion in the HAI or HIG test. Eighteen female probands, showing no serological antibody response on two occasions, by the above test methods after vaccination, demonstrated seroimmunity before vaccination documented by neutralising antibodies against rubella virus in serum samples, or a booster effect induced by the vaccination. The reason for these apparent vaccination failures could be a residual immunity following either rubella infection in utero or in earliest childhood.

Adolescent↗