Am‐3k, a Novel Monoclonal Antibody Specific for Tissue Macrophages and Its Application to Pathological Investigation

ZENG, LI; TAKEYA, Motohiro; Takahashi, Kiyoshi

doi:10.1002/(sici)1096-9896(199602)178:2<207::aid-path427>3.3.co;2-7

Cited by 14 publications

(39 citation statements)

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“…In human tissues, AM-3K reacted with most macrophages in various organs but did not react with dendritic cell populations, such as Langerhans cells, interdigitating cells, or peripheral blood monocytes (Zeng et al, 1996). In lymphoid .…”

Section: Discussionmentioning

confidence: 89%

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Interspecies reactivities of anti-human macrophage monoclonal antibodies to various animal species.

Zeng¹,

Takeya²,

Ling³

et al. 1996

J Histochem Cytochem.

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We examined interspecies reactivities of eight anti-human monocgte/maaophage monoclonal antibodies (MAbs), AM-3K, PM-2K, X4, X14, Bet-MAC3, GHIl61, EBMl11, and KP1, with various animal h e s including rats, guinea pigs, rabbits, cats, dogs, goats, pigs, M e s , horses, andmonkeys. All MAbs recognized monkey macrophages. Pig macrophages were detected by most MAbs except for EBMlll and KP1. Ofthe eight antibodies, AM3K showed the widest intespecies reactivity. It reacted with macrophages of all animal species examined, except for rats. Westem blot analysis revealed a similarity in the antigens recognized by AM-3K among guinea pigs, rabbits, and humans. Other anti-human MAbs demonstrated distinct reactive patterns against mac-

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Section: Discussionmentioning

confidence: 89%

“…Six pre-CD anti-human monocytelmacrophage MAbs, AM-3K (Zeng et al, 1996). PM-2K (Tikeya et al, 1991), X4 and X14 ( Xu et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

Interspecies reactivities of anti-human macrophage monoclonal antibodies to various animal species.

Zeng¹,

Takeya²,

Ling³

et al. 1996

J Histochem Cytochem.

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“…Fixed, paraffin-blocked tissues were sectioned serially at 2-4 µm. These were stained with routine reagents or deparaffinated, and then hydrated in PBS and treated alternatively with the AM-3K anti-monocyte/macrophage mAb (30) and VLA-5-specific antibodies.…”

Section: Methodsmentioning

confidence: 99%

Fibronectin fragments modulate monocyte VLA-5 expression and monocyte migration

et al. 1999

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To identify the mechanisms that cause monocyte localization in infarcted myocardium, we studied the impact of ischemia-reperfusion injury on the surface expression and function of the monocyte fibronectin (FN) receptor VLA-5 (α 5 β 1 integrin, CD49e/CD29). Myocardial infarction was associated with the release of FN fragments into cardiac extracellular fluids. Incubating monocytes with postreperfusion cardiac lymph that contained these FN fragments selectively reduced expression of VLA-5, an effect suppressed by specific immunoadsorption of the fragments. Treating monocytes with purified, 120-kDa cell-binding FN fragments (FN120) likewise decreased VLA-5 expression, and did so by inducing a serine proteinase-dependent proteolysis of this β 1 integrin. We postulated that changes in VLA-5 expression, which were induced by interactions with cell-binding FN fragments, may alter monocyte migration into tissue FN, a prominent component of the cardiac extracellular matrix. Support for this hypothesis came from experiments showing that FN120 treatment significantly reduced both spontaneous and MCP-1-induced monocyte migration on an FN-impregnated collagen matrix. In vivo, it is likely that contact with cell-binding FN fragments also modulates VLA-5/FN adhesive interactions, and this causes monocytes to accumulate at sites where the fragment concentration is sufficient to ensure proteolytic degradation of VLA-5.

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“…14 The antibody recognizes the first 131 amino acids residues in N-terminal region of the human ACAT-1. To confirm the formation of mature macrophages derived from cultured peripheral monocytes, we used a mouse anti-human macrophage monoclonal antibody, AM-3K, 21 generated in our laboratory. To detect rough endoplasmic reticulum by immunofluorescent and immunoelectron microscopy, we used a goat polyclonal antibody N-20 raised against a human 78-kd glucose-regulated protein, GRP 78 (Santa Cruz Biotechnology, Santa Cruz, CA).…”

Section: Antibodiesmentioning

confidence: 99%

“…21 Briefly, cultured macrophages with or without AcLDL treatment were fixed with 4% periodate-lysine-paraformaldehyde and 0.1% glutaraldehyde (Nacalai Tesque, Kyoto, Japan) in phosphate-buffered saline, pH 7.2, at 4°C for 20 minutes. After washing with phosphate-buffered saline and treating with 0.005% saponin containing phosphate-buffered saline for 10 minutes, the cells were stained by the immunoperoxidase method, using DM10 or N-20 as primary antibody.…”

Section: Immunoelectron Microscopymentioning

confidence: 99%

Localization of Human Acyl-Coenzyme A:Cholesterol Acyltransferase-1 (ACAT-1) in Macrophages and in Various Tissues

Sakashita

Miyazaki

Takeya

et al. 2000

The American Journal of Pathology

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121

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To investigate the distribution of acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT-1) in various human tissues , we examined tissues of autopsy cases immunohistochemically. ACAT-1 was demonstrated in macrophages , antigen-presenting cells , steroid hormone-producing cells , neurons , cardiomyocytes, smooth muscle cells , mesothelial cells , epithelial cells of the urinary tracts , thyroid follicles , renal tubules , pituitary , prostatic , and bronchial glands, alveolar and intestinal epithelial cells , pancreatic acinar cells , and hepatocytes. These findings showed that ACAT-1 is present in a variety of human tissues examined. The immunoreactivities are particularly prominent in the macrophages , steroid hormoneproducing cells , followed by hepatocytes , and intestinal epithelia. In cultured human macrophages, immunoelectron microscopy revealed that ACAT-1 was located mainly in the tubular rough endoplasmic reticulum; immunoblot analysis showed that the ACAT-1 protein content did not change with or without cholesterol loading; however , on cholesterol loading , about 30 to 40% of the total immunoreactivity appeared in small-sized vesicles. These vesicles were also enriched in 78-kd glucose-regulated protein (GRP 78), a specific marker for the endoplasmic reticulum. Immunofluorescent microscopy demon- Acyl-coenzyme A:cholesterol acyltransferase (ACAT) is a key enzyme involved in cellular cholesterol metabolism. It catalyzes the formation of cholesteryl esters from cholesterol and long-chain fatty acyl-coenzyme A. 1 ACAT activities are present in various tissues such as liver, intestines, adrenal glands, and aorta and are involved in intracellular cholesterol storage, lipoprotein assembly, steroid hormone production, and dietary cholesterol absorption. 2 Previous studies showed that ACAT is a membrane-bound enzyme; its activity is found only in the membrane fractions of intracellular organelles, especially in the rough endoplasmic reticulum. 3,4 More recently, molecular probes of ACAT have become available. In 1993, the first cDNA of ACAT, designated as ACAT-1, was cloned from a human THP-1 cell cDNA library by using a somatic cell and molecular genetic approach. 5 The sequence of human ACAT-1 cDNA led to the clonings of its homologues from various other species, including mice. 6 ACAT-1 gene knockout mice were produced. 7,8 Analyses of these mice showed that ACAT activities were significantly decreased in selected tissues examined but not in the livers, strongly suggesting that one or more additional ACAT genes, distinct from the ACAT-1 gene, probably exist in mice. More recently, a different ACAT cDNA, designated ACAT-2, was cloned. 9 -11 The sequence of ACAT-2 is homologous but different from that of ACAT-1. Earlier, two different ACAT-like genes were found in the simple eukaryote Sachromycetes saravesea. 12,13 The exact roles of ACAT-1 and ACAT-2 in different species remain unknown. Their physiological functions are currently under intensive investigation in several laboratories.Recently, specific polycl...

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Am‐3k, a Novel Monoclonal Antibody Specific for Tissue Macrophages and Its Application to Pathological Investigation

Cited by 14 publications

References 0 publications

Interspecies reactivities of anti-human macrophage monoclonal antibodies to various animal species.

Interspecies reactivities of anti-human macrophage monoclonal antibodies to various animal species.

Fibronectin fragments modulate monocyte VLA-5 expression and monocyte migration

Localization of Human Acyl-Coenzyme A:Cholesterol Acyltransferase-1 (ACAT-1) in Macrophages and in Various Tissues

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