Search PubMed⌕ Search

Biomedical subjects

J H Phillips

Publications and source records attributed to J H Phillips.

At least 55 records · Page 3Linked to original sources

CD94/NKG2 is the predominant inhibitory receptor involved in recognition of HLA-G by decidual and peripheral blood NK cells.

Prior studies demonstrated that NK cells isolated from adult peripheral blood kill the HLA-A-, HLA-B-, and HLA-C-deficient B lymphoblastoid cell line 721.221, but many are unable to kill 721.221 cells transfected with HLA-G, a molecule expressed preferentially on fetal cytotrophoblasts. To determine the biologic relevance of this recognition, we established NK cell clones from the placenta and demonstrate that these NK cells also were unable to kill 721.221 cells expressing HLA-G. Recognition of HLA-G by NK cells was prevented in the presence of anti-CD94 mAb, implicating CD94/NKG2 as the predominant inhibitory NK cell receptor for HLA-G used by decidual NK cells. In contrast, mAbs against the killer cell inhibitory receptors recognizing HLA-Cw3-related, HLA-Cw4-related, or HLA-Bw4 ligands did not affect NK cell killing of the HLA-G transfectants.

Adult↗

Fetal liver contains committed NK progenitors, but is not a site for development of CD34+ cells into T cells.

The presence of T and NK cells in the human fetal liver and the fact that fetal liver hemopoietic progenitor cells develop into T and NK cells suggest a role for the fetal liver compartment in T and NK cell development. In this work, we show that the capacity of fetal liver progenitors to develop into T cells, in a human/mouse fetal thymic organ culture system, is restricted to an immature subset of CD34+ CD38- cells. No T cell-committed precursors are contained within the more differentiated CD34+ CD38+ population. This conclusion is supported by the observations that no TCR-delta gene rearrangements and no pre-TCR-alpha expression can be detected in this population. However, NK cells were derived from CD34+ CD38- and CD34+ CD38+ fetal liver cells cultured in the presence of IL-15, IL-7, and Flt-3 ligand. Eighty to ninety percent of cells arising from the CD34+ CD38+ population expressed the NK cell-associated markers CD56, CD16, CD94, and NKR-P1A. Several subpopulations of NK cell precursors were identified by differential expression of these receptors. Based on the detection of populations with a similar antigenic profile in freshly isolated fetal liver cells, we propose a model of NK cell differentiation. Collectively, our findings suggest that CD34+ cells differentiate into NK cells, but not into mature T cells, in the human fetal liver.

Animals↗

Conserved and variable residues within the Bw4 motif of HLA-B make separable contributions to recognition by the NKB1 killer cell-inhibitory receptor.

Allotypes from four divergent HLA-B families (B8, B15, B16, and B27) were compared for their inhibition of cytolysis by NK cells expressing the NKB1 receptor. Allotypes differing solely at the Bw4/Bw6 region were examined as were a more divergent subset of B15 allotypes. The capacity to interact with NKB1 correlated precisely with possession of a Bw4 sequence motif at residues 77-83, whereas no correlation was made with the peptide-binding specificities of two Bw4 and four Bw6 allotypes of the B15 family. HLA-B allotypes having four different Bw4 motifs were examined and all interact with NKB1. In contrast, HLA-A allotypes, which have a Bw4 motif identical with one of those present in HLA-B, do not. Mutation at leucine 82 and arginine 83, the residues common to Bw4 motifs, shows they contribute to NKB1 interaction but are not essential. Three types of polymorphism are implicated in formation of the ligand recognized by NKB1: ones shared by Bw4 motifs; ones distinguishing Bw4 motifs; and ones outside the Bw4/Bw6 region that distinguish HLA-B from HLA-A.

Binding Sites↗

Natural killer cell cytolytic activity is inhibited by NKG2-A and activated by NKG2-C.

NKG2 is a small family of type II transmembrane proteins possessing extracellular C-type lectin domains expressed primarily on NK cells. The function of these proteins is unknown. We have developed mAbs that recognize NKG2 family members on Western blots. Examination of cell extracts from NK cell clones demonstrates that individual NKG2 family members are expressed on subsets of NK cells. Because the anti-NKG2 mAb do not react with the cell surface Ag, signaling function was studied by generating cell lines that express a chimeric receptor consisting of a cytoplasmic and transmembrane domain from NKG2-A or NKG2-C fused to the extracellular segment of mouse NKR-P1C (NK1.1 Ag). Transfectants of the rat NK cell line, RNK-16, were tested for the effect of anti-NK1.1 mAb on cytolytic activity against the FcR+ target cell lines, P388D1 and P815. The anti-NK1.1 mAb stimulated lytic activity and calcium mobilization in the NK cell line expressing the NKG2-C/NKR-P1C chimeric receptor. By contrast, anti-NK1.1 inhibited the lytic activity and failed to stimulate a calcium response in cells expressing the NKG2-A/NKR-P1C chimeric receptor. Immunoprecipitation experiments demonstrated that the inhibitory cytoplasmic tyrosine phosphatase SHP-1 selectively associates with the NKG2-A/NKR-P1C chimeric receptor, but not with the stimulatory NKG2-C/NKR-P1C receptor. These data indicate that NKG2-A and NKG2-C deliver, respectively, inhibitory and activating transmembrane signals in NK cells.

Animals↗

Functionally and structurally distinct NK cell receptor repertoires in the peripheral blood of two human donors.

The expression of KIR and CD94:NKG2 receptors was determined for more than 100 natural killer (NK) cell clones obtained from two blood donors who differ in their HLA class I and KIR genes. More than 98% of the clones were inhibited by individual autologous class I allotypes, and every clone was inhibited by the combination of autologous allotypes. The patterns of inhibition correlate with expression of inhibitory receptors of defined specificity. One donor possesses three class I ligands for KIR, and a majority of NK cells use KIR as their inhibitory receptor; the second donor possesses only a single ligand for KIR, and a majority of NK cells use the more broadly reactive CD94:NKG2a as their inhibitory receptor. Because of these differences, the first donor has subpopulations of NK cells that kill cells of the second donor, whereas the NK cells of the second donor are universally tolerant of cells from the first donor.

Antigens, CD↗

LAIR-1, a novel inhibitory receptor expressed on human mononuclear leukocytes.

In the present study, we describe a novel inhibitory receptor, leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1), that is constitutively expressed on the majority of human peripheral blood mononuclear leukocytes. LAIR-1 is a 32 kDa transmembrane glycoprotein with a single immunoglobulin-like domain and a cytoplasmic tail containing two immune receptor tyrosine-based inhibitory motifs. LAIR-1 recruits SHP-1 and SHP-2 phosphatases upon activation, and cross-linking of the LAIR-1 antigen on natural killer (NK) cells results in strong inhibition of NK cell-mediated cytotoxicity. Although it is structurally related to human killer cell inhibitory receptors, LAIR-1 does not appear to recognize human leukocyte antigen (HLA) class I molecules and thus represents a novel HLA class I-independent mechanism of NK cell regulation.

Amino Acid Sequence↗

Clinical features of Crouzon's syndrome patients with and without a positive family history of Crouzon's syndrome.

Crouzon's syndrome occurs in 1 in 25,000 live births and follows an autosomal dominant mode of transmission. However, 30 to 60% of cases are sporadic and represent fresh mutations. Previous reports involving large series of Crouzon's syndrome patients mixed sporadic and familial cases. In this article, the clinical features of 17 familial cases of Crouzon's syndrome were compared with another 27 sporadic cases. Furthermore, familial cases were studied to document (1) expressivity in members of the same family; (2) the skull base angle in unoperated members of the same family, and (3) the presence of germinal mosaicism. In familial cases of Crouzon's syndrome, craniosynostosis and proptosis were seen in 76% and 88% of patients, respectively. On the other hand, these two features were observed in 100% of sporadic cases. Variability of expression in members of the same family was a common finding. The cranial base angle was also variable in the affected members of the same family. In the series, germinal mosaicism was suspected in one family. Possible explanations for our findings are discussed as well as the implications of genetic mapping in Crouzon's syndrome.

Cephalometry↗

Design and development of a fascioperiosteal flap for use in experimental cranial surgery in a porcine model.

As part of an ongoing program of research on reconstructive techniques applicable to the growing craniofacial skeleton, this study aimed to characterize a pedicled fasciocutaneous flap on the scalp of a porcine model. Twelve juvenile Yorkshire white pigs were used. The anatomy of the porcine scalp was determined by fresh cadaveric dissection, radiocontrast dye injection, and resin casting of the vascular supply. No direct analogue of the human superficial temporal artery, the key to the superficial and deep temporal fascial flaps in humans, could be demonstrated in the pig. Thus, a fascioperiosteal flap, pedicled on an occipital leash of vessels, was designed. After a 14-day surgical delay, this flap, which covered the entire cranial width, from the nuchal crest to the glabella (7-10 cm long x 3-4 cm wide), could be elevated and split into a periosteal and a fascial layer. A cartilage construct placed between these layers could be kept alive for 4 weeks with no evidence of necrosis. The flap would allow mobilization of the construct to distant sites on the head and neck. An extensive literature survey showed a dearth of suitable cranial flap options in animal models. Accordingly, the pig cranium was examined with a view to finding a fascial flap analogous to the human temporoparietal flap, suitable for use in ear reconstruction. Cadaver dissections of the pig scalp, examining both the fascial layers and blood supply, were performed in three pigs; in an additional three pigs, the vascular supply to the scalp and cranium was injected with a radiocontrast gel, and, after dissecting suitable flaps, x-ray exposure was used to demonstrate the microvascular supply of these flaps. In an additional two pigs, the cranial vascular tree was filled with a plastic resin, and the soft tissues were then removed by thermal and chemical digestion. Finally, a group of three pigs were used to test the functional efficacy of a bilayered, occipitally based fascioperiosteal flap, whose design evolved from the foregoing work. Fluoresceine and disulphine blue were used to demonstrate the vascularity of these flaps before and after a surgical delay procedure. Function of these flaps, as carriers of a reconstructive construct, was shown by implanting autologous cartilage for 3 weeks and then demonstrating complete survival of the implant by fine section and histology. No clinically useful equivalent of the superficial temporal artery in humans could be identified in pigs, thus rendering the porcine superficial temporal artery flap of little use. However, a useful flap, comprising the pericranium and the subgaleal fascia, based on the occipital vasculature, could be raised with relative ease.

Animals↗

Arousal and inhibition of human NK cells.

NK cells express receptors for polymorphic MHC class I molecules that inhibit killing of potential target cells bearing appropriate class I allotypes. Here we review the membrane receptors on human NK cells that are known to initiate cell-mediated cytotoxicity and demonstrate regulation of these responses by the killer cell inhibitory (KIR) receptors.

Amino Acid Sequence↗

Congenital teratomas of the head and neck.

OBJECTIVE: This article reviews teratomas, neoplasms composed of the three germinal layers of the embryo that form tissues foreign to the part in which they arise. These are most common in the sacrococcygeal region and are rare in the head and neck. In this region, major concerns are airway obstruction and cosmesis. METHOD: Pathology records at two tertiary care paediatric hospitals were reviewed for diagnoses of head and neck teratomas. This revealed nine cases between 1983 and 1993, five males and four females. Two males were stillborn. Five were cervical, two were in the nasopharynx, and two were facial. The lesions were immature in two cases and mature in the remainder. All of the liveborn children underwent surgery within the first year of life, and only one required tracheotomy. RESULTS: One child had recurrence of teratomatous tissue diagnosed with CT, which was treated surgically. The child has remained free of tumour for over 3 years. Pathogenesis, clinical appearance, diagnostic work-up, and currently accepted modes of therapy are reviewed.

Female↗

Killer cell inhibitory receptor recognition of human leukocyte antigen (HLA) class I blocks formation of a pp36/PLC-gamma signaling complex in human natural killer (NK) cells.

The killer cell inhibitory receptors (KIR) of human natural killer (NK) cells recognize human leukocyte antigen class I molecules and inhibit NK cell cytotoxicity through their interaction with protein tyrosine phosphatases (PTP). Here, we report that KIR recognition of class I ligands inhibits distal signaling events and ultimately NK cell cytotoxicity by blocking the association of an adaptor protein (pp36) with phospholipase C-gamma in NK cells. In addition, we demonstrate that pp36 can serve as a substrate in vitro for the KIR-associated PTP, PTP-1C (also called SHP-1), and that recognition of class I partially disrupts tyrosine phosphorylation of NK cell proteins, providing evidence for KIR-induced phosphatase activity.

Adaptor Proteins, Signal Transducing↗

Human natural killer cell receptors involved in MHC class I recognition are disulfide-linked heterodimers of CD94 and NKG2 subunits.

CD94 receptors expressed on NK cells have been implicated in the recognition of certain HLA class I allotypes. We now demonstrate that CD94 glycoproteins form disulfide-bonded heterodimers with the NKG2A/B, NKG2C, and NKG2E glycoproteins. NKG2A/B possesses two immunoreceptor tyrosine-based inhibition motif (ITIM) sequences in its cytoplasmic domain, which may be responsible for the inhibitory function of these receptors, whereas other NKG2 proteins lack ITIMs and may potentially transmit positive signals. Structural heterogeneity in the NKG2 gene family and the formation of heterodimers with CD94 provides for the creation of a diverse NK cell repertoire.

Animals↗

The inter-locus recombinant HLA-B*4601 has high selectivity in peptide binding and functions characteristic of HLA-C.

The vast majority of new human HLA class I alleles are formed by conversions between existing alleles of the same locus. A notable exception to this rule is HLA-B*4601 formed by replacement of residues 66-76 of the alpha 1 helix of B*1501 by the homologous segment of Cw*0102. This inter-locus recombination, which brings together characteristic elements of HLA-B and HLA-C structure, is shown here to influence function dramatically. Naturally processed peptides bound by B*4601 are distinct from those of its parental allotypes B*1501 and Cw*0102 and dominated by three high abundance peptides. Such increased peptide selectivity by B*4601 is unique among HLA-A,B,C allotypes. For other aspects of function, presence of the small segment of HLA-C-derived sequence in an otherwise HLA-B framework converts B*4601 to an HLA-C-like molecule. Alloreactive cytotoxic T lymphocytes (CTL), natural killer (NK) cells, and cellular glycosidases all recognize B*4601 as though it were an HLA-C allotype. These unusual properties are those of an allotype which has frequencies as high as 20% in south east Asian populations and is associated with predisposition to autoimmune diseases and nasopharyngeal carcinoma.

Amino Acid Sequence↗

Regulation of T cell lymphokine production by killer cell inhibitory receptor recognition of self HLA class I alleles.

The killer cell inhibitory receptors (KIRs) are surface glycoproteins expressed by natural killer (NK) and T cells that specifically recognize defined groups of polymorphic human histocompatibility leukocyte antigen (HLA) class I molecules. Interactions between KIRs on NK or T cells and major histocompatibility complex (MHC) class I molecules on potential target cells inhibit cell-mediated cytotoxicity, presumably by delivering a negative signal preventing lymphocyte activation. In this study we examined whether KIRs also regulate cytokine production induced in response to T cell receptor-dependent T cell activation. CD4+ and CD8+ T cell clones were stimulated by bacterial superantigens in the presence or absence of monoclonal antibodies (mAbs) against the KIR NKB1 or MHC class I molecules, and production of tumor necrosis factor alpha and interferon gamma was evaluated. When bacterial superantigen was presented by an autologous antigen-presenting cell (APC) to a KIR+ T cell clone, cytokine production was always enhanced in the presence of anti-MHC class I mAb. Similarly, anti-KIR mAb also augmented cytokine production, provided that the APC expressed a HLA class I allele recognized by the KIR. These results suggest that recognition of autologous MHC class I molecules by KIR+ T cells provides a regulatory mechanism acting to modulate the potency of their responses to antigenic challenge.

Cells, Cultured↗

Unusual uniformity of the N-linked oligosaccharides of HLA-A, -B, and -C glycoproteins.

MHC class I glycoproteins possess an invariant site for N-linked oligosaccharide addition at position 86 of the heavy chain. For human HLA-A, -B, and -C class I glycoproteins, position 86 is the only site of N-linked glycosylation. Comparison of the size and relative abundance of oligosaccharides associated with nine HLA-A, -B, or -C allotypes isolated from EBV-transformed B cell lines and mixtures of HLA-A, -B, and -C allotypes isolated from pooled PBLs revealed a very restricted set of structures. Allotypes encoded by the HLA-A and -B loci have two predominant glycan structures that were almost exclusively di-sialylated. In contrast, HLA-C allotypes have four glycan structures, comprising those associated with HLA-A and -B and two additional glycans. Identical oligosaccharides were present on different allotypes of a class I HLA locus, and in particular, HLA-C allotypes defining two inhibitory specificities for NK cells were shown to possess the same set of oligosaccharides. The uniformity of oligosaccharide structure associated with different HLA-A, -B, and -C products and the relative lack of heterogeneity for any given allotype are unusual features for a mammalian glycoprotein. Particularly striking is that such conserved oligosaccharide structures juxtapose the major regions of amino acid sequence variation within the Ag recognition site, including the polymorphisms of the alpha 1 helix that determine the inhibitory ligands for human NK cells.

Adult↗

Inhibitory MHC class I receptors on NK cells and T cells.

Natural killer (NK) and T cells express receptors for polymorphic major histocompatibility complex (MHC) class I molecules, and activation of these receptors results in inhibition of their effector cell function. Genes encoding the murine and human NK-cell receptors have been cloned, and studies of their structure, function and expression are progressing rapidly, as reviewed here by Lewis Lanier and Joseph Phillips.

Animals↗

DNAM-1, a novel adhesion molecule involved in the cytolytic function of T lymphocytes.

Intercellular adhesion molecules play an important role in the generation of T lymphocyte-mediated immune responses. Here, we describe a novel accessory molecule, DNAX accessory molecule-1 (DNAM-1), that is constitutively expressed on the majority of peripheral blood T lymphocytes. DNAM-1 is a 65 kDa transmembrane glycoprotein consisting of 318 aa including two immunoglobulin-like domains. Anti-DNAM-1 monoclonal antibody (MAb) inhibits T and NK cell-mediated cytotoxicity against a variety of tumor cell targets and blocks cytokine production by alloantigen-specific T cells. In addition, DNAM-1 is a tyrosine-phosphorylated signal-transducing molecule that participates in primary adhesion during cytotoxic T lymphocyte (CTL)-mediated cytotoxicity.

Amino Acid Sequence↗

CD94 and a novel associated protein (94AP) form a NK cell receptor involved in the recognition of HLA-A, HLA-B, and HLA-C allotypes.

Whereas the human killer cell inhibitory receptors (KIRs) for HLA class I are immunoglobulin-like monomeric type I glycoproteins, the murine Ly49 receptors for H-2 are type II homodimers of the C-type lectin superfamily. Here, we demonstrate that human NK cells also express C-type lectin receptors that influence recognition of polymorphic HLA-A, HLA-B, and HLA-C molecules. These receptors are heterodimers composed of CD94 chains covalently associated with novel tyrosine-phosphorylated glycoproteins (94AP). Some NK clones recognize a common HLA-C ligand using both KIRs and CD94-94AP receptors. These findings suggest the existence of human inhibitory MHC class I receptors of the immunoglobulin and C-type lectin superfamilies and indicate overlap in ligand specificity.

Adult↗