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A clinical analysis of gelatin allergy and determination of its causal relationship to the previous administration of gelatin-containing acellular pertussis vaccine combined with diphtheria and tetanus toxoids.

BACKGROUND: The number of patients with allergic reactions after administration of gelatin-containing live vaccines is increasingly reported in Japan. These allergic reactions appear to be caused by gelatin allergy. It is still unknown how the patients were sensitized to gelatin. OBJECTIVE: To determine the incidence of gelatin allergy and to identify contributing factors to gelatin allergy, we investigated the following clinical aspects: the development of IgE antibodies to gelatin and the relationship of the patients' past history of acellular pertussis vaccine combined with diphtheria and tetanus toxoid (DTaP) to the development of gelatin allergy. METHODS: We evaluated 366 patient reports, submitted from 1994 to 1997, of adverse reactions after immunization with monovalent measles, mumps, and rubella vaccines containing 0.2% gelatin as stabilizer. On the basis of physician reports, the patients were categorized as to the nature of the adverse reaction. We determined the presence of IgE antibodies to gelatin and obtained past immunization history. RESULTS: The 366 reported patients were categorized as follows: 34 with anaphylaxis, 76 with urticaria, 215 with nonurticarial generalized eruption, and 41 with local reactions only. In 206 patients from whom serum was available, IgE antibodies to gelatin were detected in 25 of 27 (93%) with anaphylaxis, 27 of 48 (56%) with urticaria, and 8 of 90 (9%) with a generalized eruption. None of a group of 41 patients with only local reactions at the injected site and none of a control group of 29 subjects with no adverse reaction had such antibodies. Among 202 patients for whom prior vaccine information was available, all had received DTaP vaccines. Among those for whom the prior DTaP vaccine could be determined to contain gelatin or be free of gelatin, 155 of 158 (98%) subjects had received gelatin-containing DTaP vaccines. This rate is higher than would be expected on the basis of the market share of gelatin-containing (vs gelatin-free) DTaP vaccines (75%). Furthermore, before 1993, when a trivalent measles, mumps, and rubella vaccine (with the same 0.2% gelatin content as the monovalent vaccines) was used and administered before DTaP vaccination, no reports of anaphylaxis to the measles, mumps, and rubella vaccine were received. CONCLUSION: Most anaphylactic reactions and some urticarial reactions to gelatin-containing measles, mumps, and rubella monovalent vaccines are associated with IgE-mediated gelatin allergy. DTaP immunization histories suggest that the gelatin-containing DTaP vaccine may have a causal relationship to the development of this gelatin allergy.

Diphtheria-Tetanus-Pertussis Vaccine↗

Reactivity of the immunoglobulin E in bovine gelatin-sensitive children to gelatins from various animals.

It has been reported that most children who showed anaphylaxis to measles, mumps and rubella vaccines containing bovine gelatin as a stabilizer have anti-bovine gelatin IgE. The present study was designed to investigate the reactivity of IgE in bovine gelatin-sensitive children to gelatins from various animals, and the antigenic cross-reactivity between the gelatins. Serum samples taken from 10 children who showed anaphylaxis to vaccines containing bovine gelatin were used in this study. The level of anti-bovine gelatin IgE in these serum samples ranged from 11.0 to 251 Ua/ml. The IgE in most of the children reacted to kangaroo and mouse gelatins, to which they had had little or no exposure as a food or a vaccine stabilizer. The IgE binding to kangaroo and mouse gelatins was completely inhibited by bovine gelatin, whereas reciprocal inhibition was not complete, indicating that antigenic cross-reactivity is present between the mammalian gelatins. Only one child had strong IgE reactivity to fish gelatins, and this reactivity was not inhibited by bovine gelatin, indicating that no antigenic cross-reactivity exists between bovine and fish gelatins. Most of the children who displayed sensitivity to bovine gelatin showed IgE reactivity to other mammalian gelatins. This reactivity may be due primarily to the antigenic cross-reactivity between mammalian gelatins.

Anaphylaxis↗

Gelatin-specific cellular immune responses persist for more than 3 years after priming with gelatin containing DTaP vaccine.

BACKGROUND: Gelatin-specific cell-mediated immunity develops in subjects inoculated with gelatin containing DTaP vaccine. However, it is not yet known whether such established sensitization to gelatin disappears or persists with time. OBJECTIVE: The aim of this study was to follow the patients with gelatin sensitization elicited by DTaP vaccination for their lymphocyte responsiveness and IgE, IgG antibody specific to gelatin over several years and to compare the activities with those at the time of enrollment into the study. METHODS: We studied 28 subjects who developed positive lymphocyte proliferation test (LPT) after receiving gelatin containing DTaP vaccine and eight subjects who had a negative LPT after inoculation of non-gelatin DTaP. Determination of IgE, IgG antibodies and specific lymphoproliferative response directed against gelatin were performed at enrollment and on follow up. RESULTS: None of the subjects had antibody to gelatin at enrollment and none developed gelatin IgE or IgG during follow-up. There was no significant difference in the SIs of the subjects receiving gelatin DTaP (P = 0.150, 95% CI, -0.198-0.032), whereas lymphocyte activity to gelatin increased between enrollment and follow-up in the subjects with non-gelatin DTaP (P = 0.011, 95% CI, 0.063-0.338). CONCLUSION: Gelatin-specific lymphocyte activity persists at comparable levels for more than 3 years in subjects who acquire a positive LPT response to gelatin after receiving primary DTaP vaccine containing gelatin. Furthermore, five out of eight subjects initially with negative LPT to gelatin have been shown to acquire specific LPT with time.

Diphtheria-Tetanus-acellular Pertussis Vaccines↗

The lymphoproliferative response to enzymatically digested gelatin in subjects with gelatin hypersensitivity.

BACKGROUND: This study was designed to evaluate the immunogenic characteristics of enzymatically digested gelatin, 'FreAlagin', employing the lymphoproliferative response in subjects with gelatin hypersensitivity. Our purpose was to assess the response of primed lymphocytes to the newly developed FreAlagin and compare it to the response to conventional gelatin. METHODS: A gelatin-specific lymphocyte proliferation test (LPT) was performed in 110 children with adverse reactions to gelatin-containing vaccines, who showed positive gelatin-specific cell-mediated immunity and were thus diagnosed as having gelatin hypersensitivity. Gelatin-specific IgE was measured in all subjects. The antigenic activity of FreAlagin to lymphocytes was compared with that of conventional bovine gelatin. Positive and negative control specimens were obtained from the patients with anaphylaxis and from subjects inoculated with gelatin-free vaccine who showed no adverse reactions in order to establish the fluorometric ELISA system to determine IgE antibody to gelatin and LPT. RESULTS: The lymphocyte activity against FreAlagin was much less than that to Wako gelatin and more than half of the subjects who reacted positively to Wako gelatin had a negative LPT to FreAlagin. Although 47% of the subjects had positive LPTs to FreAlagin, all but two still had lower SIs to FreAlagin compared with Wako gelatin. CONCLUSION: We conclude that the antigenic activity of FreAlagin as measured by the cell-mediated immune response is significantly less than that of conventional bovine gelatin. However, it is still necessary to perform clinical trials to show a reduced or absent clinical reactivity to FreAlagin in sensitized patients to conventional gelatin.

Animals↗

IgE sensitization to gelatin: the probable role of gelatin-containing diphtheria-tetanus-acellular pertussis (DTaP) vaccines.

We recently found that most events of anaphylaxis to live attenuated viral vaccines containing gelatin as a stabilizer might be caused by the gelatin. However, the mechanism that the children were sensitized to gelatin was unclear. In Japan, both diphtheria-tetanus-acellular pertussis (DTaP) vaccines with and without gelatin are available. We explored the possibility that gelatin-containing DTaP vaccines before live viral vaccines sensitize children to gelatin. We received the serum samples of 87 children who had systemic immediate-type reactions including anaphylaxis to the vaccines from both physicians and vaccine manufacturers throughout Japan. We then surveyed the DTaP vaccination histories of the children who demonstrated anti-gelatin IgE. Of the above 87 children, 79 (91%) had anti-gelatin IgE. We successfully collected DTaP vaccination histories including the manufacturers' names and numbers of doses on 55 children. Only one child had not received any DTaP vaccine, the other 54 had received gelatin-containing DTaP vaccines and none received gelatin-free DTaP vaccines. We concluded that there was a strong causal relationship between gelatin-containing DTaP vaccination, anti-gelatin IgE production, and risk of anaphylaxis following subsequent immunization with live viral vaccines which contain a larger amount of gelatin.

Anaphylaxis↗

Gelatin-induced T-cell activation in children with nonanaphylactic-type reactions to vaccines containing gelatin.

BACKGROUND: Many cases of anaphylactic or nonanaphylactic reactions have been reported to measles-mumps-rubella vaccine or its component vaccines that contain gelatin as a stabilizer. Increased levels of specific IgE antibodies to gelatin have been reported in children with anaphylactic reactions. However, IgE is not increased in cases of nonanaphylactic reaction, and the mechanisms of the reaction are still controversial. OBJECTIVE: The study was aimed to elucidate the relationship between nonanaphylactic reaction and gelatin. METHODS: We investigated in vitro induction of activated memory helper T cells (CD4(+ )CD25(+ )CD45RO+ cells) in response to gelatin in children with nonanaphylactic reactions to vaccines containing gelatin. RESULTS: In patients with delayed-type sensitivity to gelatin confirmed with a positive skin test response, CD4(+ )CD25(+ )CD45RO+ cells were significantly more strongly induced in culture containing gelatin than in control cultures. However, there was no significant difference between cultures with gelatin and those with control solvent in patients without reactions after vaccination. Of 76 patients with nonanaphylactic reactions after immunization with vaccine containing gelatin, 61 had an increased lymphocyte stimulation index to gelatin versus control children. CONCLUSION: These results suggest the possibility that nonanaphylactic reactions to gelatin-containing vaccine in Japan might be mediated by delayed hypersensitivity reactions against gelatin.

Anaphylaxis↗

Physical properties of shark gelatin compared with pig gelatin.

Physical properties of shark gelatin were examined during gel formation and postgelation in comparison with pig gelatin. Samples with various concentrations and pH values were evaluated by breaking strength, dynamic viscoelasticity, and dynamic light scattering. Sol-gel and gel-sol transition temperatures for shark gelatin were remarkably lower than those for pig gelatin. Shark gelatin gel shows a narrower pH range to form a stable gel compared with pig gelatin. Melting enthalpy of shark gelatin gel was greater than that of pig gelatin gel, and G' of shark gelatin gel changed more extensively with rising temperature in comparison with pig gelatin gel. It is concluded that shark gelatin has different characteristics from pig gelatin not only for gel characteristics but also for the solution property.

Animals↗

Study on hydrophilic properties of gelatin as a clinical wound dressing. II. Water-absorbing property and hemostatic effect of gelatin.

For clinical use of gelatin as a dressing, we determined the water-absorbing capacity of gelatin for isotonic sodium chloride solution (ISCS) and bovine serum albumin-isotonic sodium chloride solution (BSA-ISCS). There was no significant difference between water absorption of gelatin with ISCS and with BSA-ISCS. The water-absorbing capacity of gelatin increased with increasing viscosity: gelatins No. 1 > No. 2 > No. 3. Enormous spaces of 20 to 30 microns or approximately 10 microns in size were observed on the gelatin surface. Scaly materials of 10 to 20 microns in size were also observed. The pores of 10 A in radius were predominant in all the gelatins studied. Since the sizes of cellular blood components range between 2 and 17 microns, they can enter the spaces but not the pores of 10 A in radius. On the other hand, water molecules, the major component of plasma, can enter the pores. Of theoretical sorption isotherms, Henderson's and Chung-Pfost's equations were fitted within the range of relative humidity between 1.5 and 75%. Gelatin No. 1 had the smallest constants n, K, B, and C followed by gelatins No. 2 and No. 3, suggesting that gelatin No. 1 is the most flexible for water and thus is fitted most easily to a raw area such as a wound or burn surface. The study of hemostatic effects of gelatin in rats demonstrated that when used at concentrations more than 50% of the bleeding amount, gelatin absorbed water from blood, and was stuck to and covered the wound surface.

Absorption↗

Interaction of anionic compounds with gelatin. II: Effect on some physicochemical properties of gelatin.

The interaction of gelatin with four monosulfonated or monocarboxylated azo dyes was investigated by measuring the surface tension and intrinsic viscosity of gelatin solutions containing the dyes at different concentrations, the rigidity and melting point of their gels, and the moisture regain of their films. The dyes, which were used as models for anionic drugs, differed in the size of their aromatic hydrocarbon moieties. Surface tension measurements showed that the gelatin did not affect the critical micelle concentration of the free dyes and that the bound dyes increased the surface activity of the gelatin. The dyes reduced the intrinsic viscosity of gelatin by as much as 2/3. They also lowered the rigidity and the melting point of dilute gelatin gels and reduced the moisture regain of dry gelatin films. These changes became more pronounced with increasing dye concentrations. The effectiveness of the dyes in producing these changes increased with the size of their hydrocarbon moieties because, as had been shown in a previous study, increasingly larger hydrocarbon moieties increased the binding of the dyes to gelatin. At the pH of the measurements, which was 1.9 units below the isoelectric point of the gelatin, the gelatin was a cationic polyelectrolyte. Binding of the dye anions by ion pairing, hydrogen bonds, and other secondary valence forces rendered the gelatin less ionic and less hydrophilic, which accounts for the present observations.

Anions↗

Gelatin-containing diphtheria-tetanus-pertussis (DTP) vaccine causes sensitization to gelatin in the recipients.

Gelatin-specific T cell response was performed to determine whether a series of vaccinations with gelatin-containing DTP is a primary sensitization process in gelatin allergy. Thirty-seven recipients with gelatin-containing DTP who developed adverse reactions after vaccination and eight recipients of DTP without gelatin who also developed adverse reactions were studied. In addition, 10 subjects receiving gelatin-containing vaccine and 10 subjects inoculated with non-gelatin vaccine who did not show any adverse reactions were also investigated. All subjects inoculated with gelatin-containing DTP vaccine showed positive T cell responses against gelatin, however, occurrence of adverse reactions did not correlate with T cell responses. We conclude that DTP vaccine containing gelatin induces sensitization to gelatin in the recipients, but the mechanism of local reactions remains unknown.

Child↗

Study on hydrophilic properties of gelatin as a clinical wound dressing. I. Hydrophilic properties of gelatin as a wound dressing.

A basic study on hydrophilicity of gelatin was made of the fitness, water-absorbing ability, water-holding ability, affinity and stability of included water in order to utilize it as a protective covering for medical use. Gelatin No. 3, being highly viscous and having a lower jelly strength, was strongly viscous and flexible, and thus, it stuck close to the surface of a lesioned site such as a wound or a burn. Since the melting point of a gelatin containing 50% water was in the range of 43 to 45 degrees C, the gelatin will be fully used as a protective covering for a wound. Below 20% relative humidity, water molecules were bound to the polar amino acid side chains of gelatin and the bound water caused a swelling of the gelatin, which is maintained by capillary sorption of water into its collagen-like four-dimensional structure. The amount of water sorbed at a relative humidity of 80% at 11 degrees C was higher by 50 to 80 mg/g compared to that at 37 degrees C. Gelatin No. 3 had a very high water-holding ability and it was very hard for it to release the sorbed water at a relative humidity of 80 to 90%. Water molecules were bound to the strong active sites of the gelatin at low relative humidity. The gelatin swelled and then, newly active sites appeared on the surface of the gelatin. The hydrophilic activity decreased with an increase in moisture content, whereas the stability of water molecules in the gelatin increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗

Contributions of the MMP-2 collagen binding domain to gelatin cleavage. Substrate binding via the collagen binding domain is required for hydrolysis of gelatin but not short peptides.

Two matrix metalloproteinases, MMP-2 and MMP-9, contain each three fibronectin type II-like modules, which form their collagen binding domains (CBDs). The contributions of CBD substrate interactions to the catalytic activities of these gelatinases have attracted special interest. Recombinant (r) CBDs retain collagen binding properties and deletions of CBDs in these MMPs reduce activities on collagen and elastin. We have characterized further the requirement of the CBD for MMP-2 cleavage of gelatin. The analyses used intact rMMP-2 and rCBD to eliminate any confounding effects that might result from structural perturbations in rMMP-2 induced by deletion of the approximately 20 kDa internal CBD. In protein-protein binding assays, 2% DMSO disrupted gelatin interactions of both rCBD and rMMP-2. At this concentration, DMSO also reduced the gelatinolytic activity by approximately 70%, pointing to a central role of CBD-substrate interactions during MMP-2 cleavage of gelatin. Subsequently, soluble rCBD was determined to competitively inhibit gelatin binding of unmodified rMMP-2 to gelatin by 73% and to reduce the MMP-2 degradation of gelatin by 70-80%. The residual gelatin cleavage that was not inhibited even by molar excess rCBD could be accounted for by degradation of short substrate molecules. Indeed, rCBD inhibited rMMP-2 cleavage of an 11 amino acid collagen-like peptide substrate (NFF-1) by less than 10%. These observations were confirmed with enzyme extracts from experimental tumors in mice. In the presence of rCBD, approximately 65% of the MMP-derived gelatinolytic activity was eliminated. Together, these results demonstrate that the CBD is absolutely required for MMP-2 cleavage of full-length collagen alpha-chains, but not for short protein fragments such as those generated by hydrolysis of gelatin.

Animals↗

IgE-mediated allergic reactions to fruit gums and investigation of cross-reactivity between gelatine and modified gelatine-containing products.

A 32-year-old female reacted with a contact urticaria syndrome after eating 'gummy bears' (fruit gums). The reaction began in the oral mucosa and led to treatment on an inpatient basis. RAST measurements with allergen discs produced with gelatine and gelatine-containing products (among them 'gummy bears') demonstrated the presence of IgE antibodies in the serum of this patient. Proteins with molecular weights in the range of 40-120 kD were identified as the allergens in gelatine by using Western blot analysis. RAST inhibition showed cross-reactivity between gelatine, gelatine-containing products and the modified gelatine used in some plasma substitutes. Allergic reactions towards coloured fruit candies and 'gummy bears' may result from an IgE-mediated hypersensitivity towards gelatine. The long-known anaphylactoid reactions towards gelatine-containing plasma substitutes may, at least in part, be of an allergic nature.

Adult↗

Acyl-gelatins for cell-hybrid biomaterials: preparation of gelatins with high melting point and affinity for hydrophobic surfaces.

In the development of cell-hybrid biomaterials, the functional activity of cells depends on the selective binding of cells to artificial ligands on the biomaterials. The extracellular matrix (ECM) is the most important ligand for cell activity. ECM is known to contain collagen, one of whose constituents is gelatin. Although natural gelatin has good cell attachment properties, the melting point of gelatin hydrogel is lower than body temperature. Thus, non-chemically cross-linked gelatin hydrogel is not a biomaterial that is used for prostheses. In the present study, we report the preparation of acyl-gelatin hydrogels with high melting point (>37 degrees C) and high affinity for hydrophobic surfaces for easy handling for transportation and adhesion activities on the hydrophobic surfaces. In addition, the doubling time of endothelial cells on the coated cell culture plate was faster than that of natural gelatin owing to the higher adhesion activity of acyl-gelatin. The results clearly demonstrated that the acyl-gelatin acted as an interface that enabled cell adhesion to artificial materials surfaces.

Acylation↗

Gelatin-binding domain-specific anti-human plasma fibronectin Fab' inhibits fibronectin-mediated gelatin binding but not cell spreading.

Antisera against a Mr = 60,000 peptide containing the gelatin-binding domain of human plasma fibronectin (McDonald, J. A., and Kelley, D. G. (1980) J. Biol. Chem. 255, 8848-8858) bound the Mr = 60,000 peptide and intact fibronectin but not three other fragments released by leukocyte elastase proteinolysis (the Mr = 25,000 amino-terminal sequence, Mr = 140,000 sequence containing cell adhesive activity, and a Mr = 31,000 fragment). Affinity-purified Fab' blocked Mr = 60,000 peptide binding to gelatin and inhibited plasma and cellular fibronectin gelatin binding without affecting fibronectin-mediated cell spreading. In contrast, antifibronectin Fab' absorbed with the gelatin-binding fragment completely blocked fibronectin-mediated cell spreading. These data indicate that the gelatin-binding domain of fibronectin is immunogenic, and antisera against this domain recognize cellular fibronectin gelatin-binding sites. Inhibition of gelatin binding but not cell spreading by anti-gelatin binding domain Fab' confirms the hypothesis that fibronectin has separate sites mediating these activities. Selective inhibition of fibronectin-collagen binding by domain-specific antisera may help elucidate the role of fibronectin in organization of the extracellular matrix.

Animals↗

Ordnance gelatin for ballistic studies. Detrimental effect of excess heat used in gelatin preparation.

Most users of ordnance gelatin for ballistics studies are apparently unaware of the detrimental effects on this tissue simulant's properties caused by excess heating in reconstitution of the gelatin powder. Material published by the Gelatin Manufacturers Institute of America states that heating gelatin above 40 degrees C can be detrimental to its properties. The manufacturer of type 250 A Ordnance Gelatin does not include directions for preparation with the gelatin powder. Directions that can be obtained by contacting the manufacturer fail to give any recommendations on the amount of heat applied during gelatin preparation and do not mention the detrimental effects of excess heat. These oversights are corrected in the revised set of directions included in this article.

Forensic Medicine↗

A novel surgical glue composed of gelatin and N-hydroxysuccinimide activated poly(L-glutamic acid): Part 1. Synthesis of activated poly(L-glutamic acid) and its gelation with gelatin.

Although fibrin glue has been widely used as a surgical adhesive, its components, fibrinogen and thrombin, obtained from human blood are not completely free from the risk of virus infection due to acquired immune deficiency and hepatitis. Recently, we have reported that a polymer pair composed of gelatin and poly(L-glutamic acid) (PLGA) promptly forms a gel and can firmly bond to soft tissues when crosslinked with the aid of water-soluble carbodiimide (WSC). The present study was undertaken to design a new PLGA-gelatin glue without using WSC. Two kinds of PLGA with molecular weights of 71 and 22 kDa were employed to prepare N-hydroxysuccinimide (NHS) activated derivatives. The NHS-activated PLGA could be synthesized at high yields and was found to be stable for an extended time without losing the ability to crosslink with gelatin when stored under a dry-cold condition. This NHS-activated PLGA could spontaneously form a gel with gelatin in an aqueous solution within a short time, comparable to a commercial fibrin glue, when gelation was allowed to proceed at pH 8.3. The NHS-activated PLGA prepared from PLGA with the molecular weight of 22 kDa could be readily dissolved at high concentrations and its ability to form a gel was maintained for more than 10 min when an acidic 8% NHS-activated PLGA solution was used. The bonding strength of PLGA gelatin glues with natural tissue was higher than that of fibrin glue. These findings strongly suggest that this combination of gelatin and NHS-PLGA is very promising as a surgical adhesive and may possibly replace fibrin glues prepared from human blood components.

Animals↗