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PubMed2002

Dissecting the gelsolin-polyphosphoinositide interaction and engineering of a polyphosphoinositide-sensitive gelsolin C-terminal half protein.

Gelsolin and other proteins in the villin/gelsolin family are regulated by polyphosphoinositides (PPIs), and manipulation of cellular PIP(2) levels alters the structure of the actin cytoskeleton coincident with the dissociation of gelsolin-actin complexes. This work explores the structure-function relationship of the gelsolin-PPI interaction. Circular dichroism experiments show that upon binding to PPIs, the PPI-sensitive N-terminal half of gelsolin undergoes significant secondary and tertiary structural changes that do not occur in the structurally homologous but PPI-insensitive C-terminal half. Secondary structure modeling algorithms predict an alpha-helical conformation for one of the gelsolin PPI-binding sites, P2, which differs from the conformation of P2 in the structure of gelsolin determined by X-ray crystallography, whereas structure prediction of the C-terminal homolog of P2 agrees well with the X-ray crystallography structure. Simulation of a change to helical conformation for P2 using molecular modeling indicates that such a structural transition will destabilize the F-actin-binding sites in domain 2. A hypothesis is proposed that PPIs initiate conformational changes at the PPI-binding site(s) that destabilize the protein structure, and subsequently disrupt the actin-binding sites. To further evaluate the role of P2 in the gelsolin-PPI interaction, a Ct mutant P2Ct is constructed by inserting P2 in place of its C-terminal homologous site. P2Ct interacts with actin in the same way as the wild-type protein. In contrast to Ct, however, P2Ct interacts strongly with PPIs, and its monomeric actin-binding activity becomes regulated by PPIs. It is concluded that the P2 site is sufficient for PPI-sensitivity in gelsolin. Furthermore, the P2 site in P2Ct and the actin-binding sites of Ct do not overlap, suggesting that PPIs regulate actin binding of P2Ct through induction of structural changes, rather than through direct competition.

Actins↗

PubMed2003

Recent advances in disc cell biology.

STUDY DESIGN: There have been many advances over the past decade in understanding and experimentally modulating biologic aspects of intervertebral disc cell function. An overview of the current state of this biologic research is presented. OBJECTIVES: To provide clinicians with a review of important recent advances in biologic studies of the disc and their implications for potential disc therapies. SUMMARY OF BACKGROUND DATA: Historically, anatomic, biochemical, radiologic, and biomechanical studies of the intervertebral disc formed the foundation on which our understanding of disc function was built. Magnetic resonance imaging techniques that allowed viewing of soft tissue components of the disc further advanced imaging capabilities. METHODS: Recent publications are reviewed. RESULTS: Experimental approaches over the past decade have enabled researchers to look more critically at disc cell function. This is important because disc cell function produces the extracellular matrix components of the disc, which, in turn, shape the disc's subsequent physiologic and biomechanical functions. New approaches to the study of disc cell function, methods to manipulate disc cells, studies of intact discs and disc nutrition, vertebral endplate structure and function, tissue engineering, gene therapy, and the potential of stem cells in disc therapy are reviewed and discussed. CONCLUSIONS: Many believe that disc degeneration has a cellular basis. New research is helping us better understand healthy, aging, and degenerating discs. Modern methods to manipulate and modulate disc cell function open exciting and challenging new therapeutic possibilities for future biologic treatments of disc degeneration.

Animals↗

PubMed2002

Adult mesenchymal stem cells and cell-based tissue engineering.

The identification of multipotential mesenchymal stem cells (MSCs) derived from adult human tissues, including bone marrow stroma and a number of connective tissues, has provided exciting prospects for cell-based tissue engineering and regeneration. This review focuses on the biology of MSCs, including their differentiation potentials in vitro and in vivo, and the application of MSCs in tissue engineering. Our current understanding of MSCs lags behind that of other stem cell types, such as hematopoietic stem cells. Future research should aim to define the cellular and molecular fingerprints of MSCs and elucidate their endogenous role(s) in normal and abnormal tissue functions.

Adult↗

PubMed2001

Toward a molecular understanding of psychostimulant actions using genetically engineered dopamine receptor knockout mice as model systems.

A major focus in studying the progression and prevention of addictive diseases has been to understand the molecular and cellular mechanisms underlying drug addiction. The brain dopaminergic system plays a central role in reward and motivation and is thought to be the main neural substrate for the actions of abusive drugs. We have used the gene targeting technology to generate dopamine D1 and D3 receptor knockout mice and used these mice as model systems to gain a molecular understanding of acute effects of psychostimulants cocaine and amphetamine. The use of a combined approach involving behavioral, electrophysiological as well as molecular studies has allowed us to define initially the roles of dopamine D1 and D3 receptors in the acute effects of psychostimulants and will enable us to understand mechanisms underlying their chronic actions in the future.

Amphetamine↗

PubMed2026

A modular γδ TCR-T platform combining KRAS pMHC targeting with re-dosable mRNA engager redirection.

Solid tumors often evade TCR-engineered αβ T cells when antigen expression varies or when the restricting Human Leukocyte Antigen (HLA) allele is lost. γδ T cells, in contrast, detect cellular dysregulation through non-peptide/Major Histocompatibility Complex (MHC) cues, including phosphoantigens and stress ligands, and can be developed as allogeneic therapies. Although intratumoral γδ T cell signatures are associated with improved outcome across cancers, γδ recognition itself is broad and still selected within the thymus just as αβ T cell receptors (TCRs) are. It does not, however, anchor specificity to a defined driver-mutation pMHC epitope. We therefore asked whether a high-affinity, co-receptor-independent αβ TCR could graft oncogenic-driver specificity onto γδ T cells while leaving the endogenous γδ TCR intact. We knocked the KRASG12V/HLA-A*11:01 TCR A11v into primary human γδ T cells. Engineered cells co-expressed the transgenic αβ TCR and the endogenous γδ TCR and lysed KRASG12V/HLA-A*11:01+ tumor cells in vitro and in vivo. To cover potential resistance through loss of HLA-A*11:01, we delivered an mRNA lipid nanoparticle (LNP) encoding a secreted mesothelin×CD3 (M5) bispecific T cell engager (TCE). LNP-M5 produced circulating TCE that redirected γδ A11v T cells and polyclonal bystander T cells to kill mesothelin+ targets, accompanied by development of higher γδ A11v T cell counts in vivo. In humanized mice bearing mixed HLA-A*11:01+ and HLA-A*11:01 - KRASG12V tumors, γδ A11v T cells produced transient control, whereas adding LNP-M5 yielded complete responses and prolonged survival. Thus, this two-part therapy couples invariant driver targeting to tunable redirection and addresses loss of the restricting HLA allele, a central escape route for TCR-based therapy. It provides an off-the-shelf reagent to enable KRAS-anchored treatment with the ability to redeliver the reagent.

Humans↗

PubMed2006

[Cartilage formation and regeneration by mechanical stress].

Every cell in our body is subject to mechanical stress and responds to mechanical stimulation. Especially, tissues which function as a supportive tissue such as bone or cartilage utilize these stimulation during tissue formation and maintenance. Several lines of extensive studies have revealed the effects and mechanisms of mechanotransduction in cartilage or three dimensional engineered tissue. Furthermore, these biological responses to mechanical stimulation are thought to enhance cartilage formation or regeneration in vitro . Mechanical stimulation to a cultured tissue is thought to be a feasible strategy to develop a new cartilage therapy.

Animals↗

PubMed2006

Cyclic flexure and laminar flow synergistically accelerate mesenchymal stem cell-mediated engineered tissue formation: Implications for engineered heart valve tissues.

Bone marrow-derived mesenchymal stem cells (BMSCs) are relatively accessible and exhibit a pluripotency suitable for cardiovascular applications such as tissue-engineered heart valves (TEHVs). Recently, Sutherland et al. [From stem cells to viable autologous semilunar heart valve. Circulation 2005; 111(21): 2783-91] demonstrated that BMSC-seeded TEHV can successfully function as pulmonary valve substitutes in juvenile sheep for at least 8 months. Toward determining appropriate mechanical stimuli for use in BMSC-seeded TEHV cultivation, we investigated the independent and coupled effects of two mechanical stimuli physiologically relevant to heart valves-cyclic flexure and laminar flow (i.e. fluid shear stress)-on BMSC-mediated tissue formation. BMSC isolated from juvenile sheep were expanded and seeded onto rectangular strips of nonwoven 50:50 blend poly(glycolic acid) (PGA) and poly(l-lactic acid) (PLLA) scaffolds. Following 4 days static culture, BMSC-seeded scaffolds were loaded into a novel flex-stretch-flow (FSF) bioreactor and incubated under static (n=12), cyclic flexure (n=12), laminar flow (avg. wall shear stress=1.1505 dyne/cm(2); n=12) and combined flex-flow (n=12) conditions for 1 (n=6) and 3 (n=6) weeks. By 3 weeks, the flex-flow group exhibited dramatically accelerated tissue formation compared with all other groups, including a 75% higher collagen content of 844+/-278 microg/g wet weight (p<0.05), and an effective stiffness (E) value of 948+/-233 kPa. Importantly, collagen and E values were not significantly different from values measured for vascular smooth muscle cell (SMC) -seeded scaffolds incubated under conditions of flexure alone [Engelmayr et al. The independent role of cyclic flexure in the early in vitro development of an engineered heart valve tissue. Biomaterials 2005; 26(2): 175-87], suggesting that BMSC-seeded TEHV can be optimized to yield results comparable to SMC-seeded TEHV. We thus demonstrated that cyclic flexure and laminar flow can synergistically accelerate BMSC-mediated tissue formation, providing a basis for the rational design of in vitro conditioning regimens for BMSC-seeded TEHV.

Animals↗

PubMed2004

New detection system for toxic agents based on continuous spectroscopic monitoring of living cells.

In this study, we show the feasibility of a new type of cell based biosensor which uses spectroscopic in situ real time detection of biochemical changes in living cells exposed to toxic chemical agents. We used a high power 785 nm laser to measure the time dependent changes in the Raman spectrum of individual living human lung cells (A549 cell line) treated with a toxic agent (Triton X-100, 250 microM solution). Individual cells were monitored by Raman spectroscopy over a total time span of 420 min, with 30 min sampling intervals. During this period of time, the A549 cells were maintained in a purpose designed temperature controlled cell chamber, which allowed the cells to be maintained in physiological conditions. The time dependent changes in the Raman spectra were correlated with the sequences of events that occur during cell death. The molecular mechanisms involved in cell death are indicated by the decrease in the magnitude of Raman peaks corresponding to proteins (1322, 1342 and 1005 cm(-1)) and DNA (decrease by 80-90% in the 786 cm(-1) phosphodiester bonds C'5-O-P-O-C'3). To support these conclusions, viability tests and Western blotting analysis of PARP protein were carried out. This technique could overcome the limitations of other detection systems available, since the specific time dependent biochemical changes in the living cells can be used for the identification and quantification of a large range of toxic agents. This technique could also be used with cellular microarrays for high throughput in vitro toxicological testing of pharmaceuticals and in situ monitoring of the growth of engineered tissues.

Adenocarcinoma↗

PubMed2006

Targeting of HIV-1 Tat traffic and function by transduction-competent single chain antibodies.

Human immunodeficiency virus type 1-encoded Tat protein is a transactivating factor essentially required for viral replication. Tat binds specifically to the transactivation response RNA stem loop, which is formed at the 5' end of all viral transcripts. The TAR binding motif of Tat also contains a protein transduction domain, PTD that mediates not only nuclear localization of Tat but is also capable of transducing cargo across cellular membranes. In order to target a Tat antagonist directly to the TAR binding site in the nucleus, we engineered a chimeric protein consisting of the Tat-derived PTD fused to the anti-Tat single chain antibody scFvtat1 that binds intracellularly to Tat. Recombinant scFvtat1-PTD(TAT) fusion antibody retained both, anti-Tat specificity and PTD(TAT)-mediated transduction-competence leading to its nuclear accumulation within living cells. Incubation of Jurkat T cells with scFvtat1-PTD(TAT) suppressed Tat-dependent transcription of a HIV-1 reporter gene by >80%. Transfection of a scFvtat1-PTD(TAT) expression plasmid in HEK293 cells resulted in diffuse cytoplasmic and nuclear expression. ScFvtat1-PTD(TAT) did not inhibit HIV-1 Tat translocation to the nucleus, yet showed increased inhibition of 78%, indicating a nuclear site of scFvtat1-PTD(TAT) action. Strikingly, the PTD(TAT) alone showed 55% inhibition in the HIV-1 luciferase reporter assay, indicating competition with HIV-1 Tat binding to the TAR element. The results of this study suggest that Tat traffic can only marginally be affected by anti-Tat antibodies and that effective inhibition of Tat function requires both competition with HIV Tat for TAR binding mediated by PTD(TAT) and steric hindrance mediated by the scFvtat1 moiety.

Amino Acid Motifs↗

PubMedDate not supplied

Modulation of in vitro myogenesis induced by different polymer substrates.

The understanding of substrate dependence of cellular differentiation is important in the surface design of biocompatible artificial devices as well as cell-incorporated tissue engineered devices. In an attempt to understand some of the genetic and epigenetic aspects of the control of cell differentiation in the presence of two different materials, Chronoflex (CH) and plasma treated Chronoflex coated with Hyaluronan (CH-HA), we used primary cultures of human myogenic cells, a model that encompasses cell proliferation, migration, fusion, and differentiation dependent gene activation. By testing both the material samples on the growth of human myoblasts in primary cultures, we demonstrated that both CH and CH-HA substrates were able to support the cell growth since they did not affect cell count and DNA synthesis. On the contrary, the degree of myoblast differentiation, assessed as a function of creatine phosphokinase (CPK) activity on living cells, was completely different on the two biomaterials. Indeed, the amount of CPK increased on CH-HA cultured cells as a result of myotube formation, while CH grown myoblasts remained unfused and displayed no increase on the CPK activity even after 12 days culture. Moreover, the expression level of MyoD and myogenin mRNA, both related to myogenic cell differentiation, appeared extremely low in CH-grown cells, while they were rapidly induced in CH-HA cultured myoblasts.

Journal Article↗

PubMed1998

Signaling pathways underlying eosinophil cell motility revealed by using caged peptides.

Insights into structure-function relations of many proteins opens the possibility of engineering peptides to selectively interfere with a protein's activity. To facilitate the use of peptides as probes of cellular processes, we have developed caged peptides whose influence on specific proteins can be suddenly and uniformly changed by near-UV light. Two peptides are described which, on photolysis of a caging moiety, block the action of calcium-calmodulin or myosin light chain kinase (MLCK). The efficacy of theses peptides is demonstrated in vitro and in vivo by determining their effect before and after photolysis on activities of isolated enzymes and cellular functions known to depend on calcium-calmodulin and MLCK. These caged peptides each were injected into motile, polarized eosinophils, and when exposed to light promptly blocked cell locomotion in a similar manner. The results indicate that the action of calcium-calmodulin and MLCK, and by inference myosin II, are required for the ameboid locomotion of these cells. This methodology provides a powerful means for assessing the role of these and other proteins in a wide range of spatio-temporally complex functions in intact living cells.

Animals↗

PubMed1999

Phthalimide analogs of CB 1954: synthesis and bioactivation.

Four novel 4-substituted 5-nitrophthalimides (5-substituted-6-nitro-1,3-dihydro-isoindol-1,3-diones), 6, 7, 10 and 11, and the known 5 are prepared as analogs of the dinitrobenzamide prodrug CB 1954, 1, and considered as potential candidates for gene-directed enzyme prodrug therapy. All the phthalimides are poor substrates for Escherichia coli nitroreductase compared to CB 1954. However, 6, 7, 10 and 11 are reduced by both the human and rat forms of DT-diaphorase; 10 is a particularly good substrate but 7 decomposes in phosphate buffer. A cell-line panel consisting of V79 cells that have been engineered to express various levels of either the human or rat forms of DT-diaphorase in an identical cellular background was used to evaluate these compounds as prodrugs activated by this enzyme. The cytotoxic effect of CB 1954 is proportional to the activity of either the rat or human enzyme but cells expressing the rat enzyme were much more sensitive (10000-fold at higher levels of DT-diaphorase activity) than cells expressing comparable levels of the human enzyme. These results demonstrate that the resistance of human tumors to CB 1954 can be accounted for solely by the kinetic properties of the enzyme for this prodrug. The nitrophthalimide analogs overcome this kinetic failing of CB 1954. However, these compounds are not activated to produce cytotoxicity in these DT-diaphorase-expressing cell lines. It is postulated their reduction products fail to undergo an acylation reaction in a manner analogous to CB 1954. Thus, reduction by DT-diaphorase is not predictive of cytotoxicity in this class of prodrugs.

Animals↗

PubMed1999

Meniscal substitutes--animal experience.

Animal studies have shown that meniscus allografts and tendon autografts generally heal to the capsule, are revascularized and repopulated with host cells. In animals, neither meniscal allografts nor tendon or fat autografts gain the properties of a normal meniscus. Meniscus allografts and tendon autografts are promising as both seem to offer some protection to the cartilage of the tibial plateau. There is no evidence that meniscal transplantation can prevent cartilage degenerative changes, and the long-term effect of meniscal transplantation on articular cartilage remains unknown. Whether cellular repopulation of the meniscal allograft is sufficient to restore its biomechanical properties is unknown. Collagen scaffolds and tissue engineered grafts are still under investigation, showing promising results especially for the former. Viable meniscal allografts should be implanted within 1 to 2 weeks after harvesting, as the production of proteoglycans decreases after 2 weeks.

Animals↗

PubMed2002

Human salivary histatin 5 causes disordered volume regulation and cell cycle arrest in Candida albicans.

Human salivary histatin 5 (Hst 5) is a nonimmune salivary protein with antifungal activity against an important human pathogen, Candida albicans. The candidacidal activity of histatins appears to be a distinctive multistep mechanism involving depletion of the C. albicans intracellular ATP content as a result of nonlytic ATP efflux. Hst 5 caused a loss of cell viability concomitant with a decrease in cellular volume as determined both by a classical candidacidal assay with exogenous Hst 5 and by using a genetically engineered C. albicans strain expressing Hst 5. Preincubation of C. albicans cells with pharmacological inhibitors of anion transport provided complete or substantial protection from Hst 5-induced killing and volume reduction of cells. Moreover, intracellular expression of Hst 5 resulted in a reduction in the population mean cell volume that was accompanied by an increase in the percentage of unbudded cells and C. albicans cells in the G(1) phase. Following expression of Hst 5, the smallest cells sorted by fluorescence-activated cell sorting from the total population did not replicate and were exclusively in the G(1) phase. Cells with intracellularly expressed Hst 5 had greatly reduced G(1) cyclin transcript levels, indicating that they arrested in the G(1) phase before the onset of Start. Our data demonstrate that a key determinant in the mechanism of Hst 5 toxicity in C. albicans cells is the disruption of regulatory circuits for cell volume homeostasis that is closely coupled with loss of intracellular ATP. This novel process of fungicidal activity by a human salivary protein has highlighted potential interactions of Hst 5 with volume regulatory mechanisms and the process of yeast cell cycle control.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1998

Requirement of cyclin E-Cdk2 inhibition in p16(INK4a)-mediated growth suppression.

Loss-of-function mutations of p16(INK4a) have been identified in a large number of human tumors. An established biochemical function of p16 is its ability to specifically inhibit cyclin D-dependent kinases in vitro, and this inhibition is believed to be the cause of the p16-mediated G1 cell cycle arrest after reintroduction of p16 into p16-deficient tumor cells. However, a mutant of Cdk4, Cdk4(N158), designed to specifically inhibit cyclin D-dependent kinases through dominant negative interference, was unable to arrest the cell cycle of the same cells (S. van den Heuvel and E. Harlow, Science 262:2050-2054, 1993). In this study, we determined functional differences between p16 and Cdk4(N158). We show that p16 and Cdk4(N158) inhibit the kinase activity of cellular cyclin D1 complexes through different mechanisms. p16 dissociated cyclin D1-Cdk4 complexes with the release of bound p27(KIP1), while Cdk4(N158) formed complexes with cyclin D1 and p27. In cells induced to overexpress p16, a higher portion of cellular p27 formed complexes with cyclin E-Cdk2, and Cdk2-associated kinase activities were correspondingly inhibited. Cells engineered to express moderately elevated levels of cyclin E became resistant to p16-mediated growth suppression. These results demonstrate that inhibition of cyclin D-dependent kinase activity may not be sufficient to cause G1 arrest in actively proliferating tumor cells. Inhibition of cyclin E-dependent kinases is required in p16-mediated growth suppression.

Animals↗

PubMed2001

Role of alternative splicing in generating isoform diversity among plasma membrane calcium pumps.

Calcium pumps of the plasma membrane (also known as plasma membrane Ca(2+)-ATPases or PMCAs) are responsible for the expulsion of Ca(2+) from the cytosol of all eukaryotic cells. Together with Na(+)/Ca(2+) exchangers, they are the major plasma membrane transport system responsible for the long-term regulation of the resting intracellular Ca(2+) concentration. Like the Ca(2+) pumps of the sarco/endoplasmic reticulum (SERCAs), which pump Ca(2+) from the cytosol into the endoplasmic reticulum, the PMCAs belong to the family of P-type primary ion transport ATPases characterized by the formation of an aspartyl phosphate intermediate during the reaction cycle. Mammalian PMCAs are encoded by four separate genes, and additional isoform variants are generated via alternative RNA splicing of the primary gene transcripts. The expression of different PMCA isoforms and splice variants is regulated in a developmental, tissue- and cell type-specific manner, suggesting that these pumps are functionally adapted to the physiological needs of particular cells and tissues. PMCAs 1 and 4 are found in virtually all tissues in the adult, whereas PMCAs 2 and 3 are primarily expressed in excitable cells of the nervous system and muscles. During mouse embryonic development, PMCA1 is ubiquitously detected from the earliest time points, and all isoforms show spatially overlapping but distinct expression patterns with dynamic temporal changes occurring during late fetal development. Alternative splicing affects two major locations in the plasma membrane Ca(2+) pump protein: the first intracellular loop and the COOH-terminal tail. These two regions correspond to major regulatory domains of the pumps. In the first cytosolic loop, the affected region is embedded between a putative G protein binding sequence and the site of phospholipid sensitivity, and in the COOH-terminal tail, splicing affects pump regulation by calmodulin, phosphorylation, and differential interaction with PDZ domain-containing anchoring and signaling proteins. Recent evidence demonstrating differential distribution, dynamic regulation of expression, and major functional differences between alternative splice variants suggests that these transporters play a more dynamic role than hitherto assumed in the spatial and temporal control of Ca(2+) signaling. The identification of mice carrying PMCA mutations that lead to diseases such as hearing loss and ataxia, as well as the corresponding phenotypes of genetically engineered PMCA "knockout" mice further support the concept of specific, nonredundant roles for each Ca(2+) pump isoform in cellular Ca(2+) regulation.

Alternative Splicing↗

PubMed1998

Structure and functional characteristics of bacterial biofilms in fluid processing operations.

Bacterial biofilms create a number of serious problems for industrial fluid processing operations. Mechanical blockages, impedance of heat transfer processes, and biodeterioration of the components of metallic and polymeric systems result in billions of dollars in losses each year. Product spoilage and possible risks to public health are also consequences of biofilm-mediated contamination. Fundamentally, these biofouling activities can be described in terms of the physicochemical properties that are associated with bacterial metabolism and biofilm development. Treatment of biofouling is also complicated by the unique structural attributes of biofilms: extracellular polymeric substances create diffusional barriers to antimicrobial agents, protecting labile cellular targets from both oxidizing and nonoxidizing compounds. The mechanisms associated with the initial events of bacterial adhesion to engineered surfaces and subsequent fouling of biofilm formation are poorly understood. However, studies of bacterial biofilm architecture have been greatly facilitated by the application of confocal laser microscopy, scanning or transmission electron microscopy, and Fourier transform infrared spectroscopy. This paper reviews the genesis of biofilm formation and describes the influence of structure on biofouling activities in industrial fluid handling systems.

Animals↗

PubMed2004

[Construction of recombinant fowlpox virus coexpressing gp120 of Chinese HIV-1 strain and IL-18 and its immunogenicity in mice].

To screening out Chinese vaccine candidate against HIV-1, Chinese vaccine strain 282E4 of fowlpox virus was used as the vector to construct the recombinant fowlpox virus (rFPV) coexpressing gp120 of Chinese HIV-1 strain and IL-18, and the recombinant virus was indentified by PCR and Western blot. The specific DNA fragment could be amplified by PCR from the genome of rFPV. Western blot analysis showed that gp120 and IL-18 could be expressed not only in chicken embryo fibroblast (CEF) cells infected by rFPV, but also in mammalian cells infected by rFPV. After the recombinant fowlpox virus was inoculated into BALB/c mice, the spleen specific CTL activities and serum antibodies in the immunized mice were detected, which demonstrated that the rFPV had good immunogenicity and could induce BALB/c mice to produce specific humoral and cellular immunity. IL-18 palyed the role of immunoadjuvant. The study lays the basis on the preparation of genetic engineering live vector vaccine against HIV-1.

AIDS Vaccines↗