Vaccine Injection Site Research Articles

Autologous Tumor Combined with a GM-CSF-Secreting Cell Line Vaccine (GVAX®) Following Autologous Stem Cell Transplant (ASCT) in Multiple Myeloma.

Background: Preclinical models have demonstrated enhanced cancer vaccine efficacy when administered early following ASCT and accompanied by vaccine-primed lymphocyte infusion.Methods: Patients with untreated myeloma were enrolled and underwent a bone marrow harvest to obtain tumor cells for vaccine processing. Patients eligible for ASCT following induction chemotherapy received a pre-transplant vaccination followed by leukapheresis to collect “primed” lymphocytes that were reinfused with the stem cell graft. Eight post-transplant vaccinations were administered at 3-week intervals starting 6 weeks posttransplant. The vaccine formulation consisted of irradiated autologous tumor cells admixed with GM-CSF-secreting K562 cells (CG9962) at a ratio of 5:2, respectively. Three dose levels were assigned (1, 4, and 10 x107 tumor cells/vaccination) based on tumor cell yield from the bone marrow harvest.Results: 22 patients underwent tumor harvest, 18 completed induction chemotherapy, and 16 proceeded to vaccination (1 at 1 x 107,4 at 4x107, and 13 at 10x107) and ASCT. At baseline, the median b2 microglobulin was 4 (2–14) and 35% had an IgA subtype. No grade 3/4 vaccine-related toxicities were observed. Median time to neutrophil engraftment (>500) and platelet engraftment (>50K) was 10 days. Local vaccine injection site reactions were observed in all patients. Delayed-type hypersensitivity (DTH) reactions to injections of irradiated autologous tumor cells were induced in only 1/15 patients after 4 post-transplant vaccines, but 4/10 were positive at one year post transplant. Induction of autologous tumor-reactive antibodies as well as antibodies reactive against CG9962 cells was observed. In vitro studies have demonstrated the induction and adoptive transfer of tumor-specific cellular immune responses that can be followed throughout the post-transplant period. Reproducible serum GM-CSF levels were achieved with repeated vaccinations suggesting no increased clearance of CG9962 cells due to induction of an allogeneic immune response. Of the 16 patients that have completed the ASCT, there were 6 complete and 5 partial remissions, for an overall response rate of 69%. Three patients with rising paraprotein levels early post-transplant have demonstrated paraprotein declines following initiation of post-transplant vaccines, suggesting a possible vaccine-mediated effect.Conclusions: These data demonstrate the feasibility of this vaccine approach and show the therapeutic potential of K562 GVAX® vaccination in the ASCT setting in multiple myeloma.

Posttransplant Immunotherapy with a GM-CSF-Based Tumor Vaccine (GVAX®) Following Autologous Stem Cell Transplant (ASCT) for Acute Myeloid Leukemia (AML).

Background: Preclinical models have demonstrated the efficacy of GM-CSF secreting cancer vaccines accompanied by vaccine-primed lymphocyte infusion following ASCT.Methods: Patients ≤ 60 years old with de novo AML (excluding M3) were enrolled. Leukemia cells were harvested at diagnosis followed by induction and consolidation chemotherapy and ASCT. A pretransplant vaccine composed of irradiated autologous leukemia cells mixed with GM-CSF gene-modified K562 cells (CG9962) was given followed by collection of vaccine-primed lymphocytes that were reinfused with the stem cell graft. Posttransplant vaccinations were initiated at week 6 (or upon platelet engraftment) and given every 3 weeks (10x107 tumor cells + 4x107 CG9962 cells) x 8 vaccinations.Results: Leukemia cell harvest was successful in 51/54 patients (blood draw − 22, leukapheresis − 28, bone marrow aspirate − 4). To date, 44/54 patients (81%) have achieved a complete remission (CR) following induction chemotherapy, 27 have received the pretransplant vaccination, and 15 have initiated posttransplant vaccinations. Local vaccine injection site reactions developed in all patients. Delayed-type hypersensitivity (DTH) reactions to irradiated, autologous tumor were induced post vaccination in 4/11. Antibodies reactive against autologous tumor were induced in 5/11 and against CG9962 cells in 10/10. Minimal residual disease (MRD) is being monitored by quantitative analysis of peripheral blood (PB) and bone marrow (BM) for WT1, a leukemia-associated gene, by RT-PCR. All patients had detectable WT1 transcripts in PB and BM at diagnosis. Most patients in CR had persistently detectable WT1 transcripts in BM following induction (92%), consolidation (91%), and ASCT (63%), whereas, clearance of WT-1 in PB was more frequent (49% detectable post induction, 50% post consolidation, and 31% post ASCT). A decrease in WT1 was noted in 67% (12/18 in the BM and 10/15 in PB) following the pretransplant vaccination.Conclusions: Collection of large quantities of autologous leukemia cells for vaccine production is feasible. Reduction in WT-1 transcript levels following the pre-transplant vaccine suggests anti-leukemic activity of this GVAX® vaccine platform.

Aluminium assay and evaluation of the local reaction at several time points after intramuscular administration of aluminium containing vaccines in the Cynomolgus monkey

Aluminium hydroxide and aluminium phosphate have been widely used as vaccine adjuvants with a good safety record for several decades. The recent observation in human deltoid muscle of macrophage aggregates containing aluminium hydroxide spicules and termed Macrophagic Myofasciitis (MMF) has encouraged research on aluminium salts. This study was conducted in order to further investigate the clearance of aluminium at the vaccine injection site and the features of induced histopathological lesions. Two groups of 12 monkeys were immunised in the quadriceps muscle with Diphtheria–Tetanus vaccines, which were adjuvanted with either aluminium hydroxide or aluminium phosphate. Three, six or twelve months after vaccination, four monkeys from each group were sacrificed and histopathological examination and aluminium assays were performed on quadriceps muscle sections. Histopathological lesions, similar to the MMF described in humans, were observed and were still present 3 months after aluminium phosphate and 12 months after aluminium hydroxide adjuvanted vaccine administration. An increase in aluminium concentration, more marked in the area of the lesions, was also observed at the 3- and 6-month time points. These findings were localised at the injection site and no similar changes were observed in the distal or proximal muscle fragments. We conclude from this study that aluminium adjuvanted vaccines administered by the intramuscular route trigger histopathological changes restricted to the area around the injection site which persist for several months but are not associated with abnormal clinical signs.

Safety and immunogenicity of a polyvalent peptide C4-V3 HIV vaccine in conjunction with IL-12.

We examined the safety and immunogenicity of a human leukocyte antigen-based HIV envelope polyvalent synthetic peptide vaccine, C4-V3, alone and in combination with subcutaneous IL-12 in nine HIV-infected patients. Lymphocyte proliferative responses increased threefold or more to all four peptides at two consecutive post-immunization timepoints for four individuals. Three responders had received IL-12, suggesting a possible adjuvant effect of Il-12. Transient mild injection site reactions (7.9) and systemic symptoms (3/9) occurred.

A DNA vaccine directed against a rainbow trout rhabdovirus induces early protection against a nodavirus challenge in turbot

A DNA vaccine encoding the envelope glycoprotein from a fish rhabdovirus, viral hemorrhagic septicemia virus (VHSV), has previously been shown to induce both early and long time protection against the virus in rainbow trout. Challenge experiments have revealed that the immunity established shortly after vaccination is cross-protective against heterologous fish rhabdoviruses. In this study, we show that the DNA vaccine encoding the VHSV glycoprotein also induces early protection against a non-enveloped, positive-sense RNA virus belonging to the Nodavirus family, the Atlantic halibut nodavirus (AHNV). In a vaccine efficacy test using juvenile turbot as model fish, the fish injected with the VHSV vaccine were completely protected against a nodavirus challenge performed 8 days post vaccination, while the cumulative mortality in the control group reached 54%. A DNA vaccine carrying the gene encoding the capsid protein of AHNV revealed no protective properties against the nodavirus challenge. Histological examination of muscle tissue sections from the vaccine injection site showed that the DNA vaccine against VHSV triggered a pronounced inflammatory response in turbot similar to what has earlier been observed in rainbow trout.

Mechanism and therapeutic potential of DNA-based immunization against the envelope proteins of hepatitis B virus in normal and transgenic mice.

Two plasmid DNA vectors, pCAGGS(S) encoding the genes of the major envelope protein of hepatitis B virus (HBV), and pCAGGS(S + preS2) encoding the genes of the middle envelope protein were used to study the mechanism and therapeutic potential of DNA-based immunization. Injection of these plasmids into the regenerating bilateral tibialis anterior muscle (TA) of normal C57BL/6 mice induced hepatitis B surface antigen (HBsAg)-specific humoral and cellular immune responses. Seventy-two hours after injection of pCAGGS(S), infiltrating cells including antigen-presenting dendritic cells (DC) were localized around the injection site and HBsAg was expressed by both muscle cells and infiltrating cells. Spleen DC from the mice were exposed to HBsAg for up to 32 weeks after a single injection of pCAGGS(S), because these DC induced the proliferation of HBsAg-specific memory lymphocytes in culture without exogenous HBsAg. A single injection of pCAGGS(S) or pCAGGS(S + preS2) resulted in the clearance of HBsAg in 28 out of 30 HBV-transgenic (Tg) mice. In contrast, more than 7 monthly injections of an HBsAg-based vaccine were required for the clearance of HBsAg in 6 out of 29 HBV-Tg mice. Infiltrating DC at the DNA vaccine injection site may have a role in initiating HBsAg-specific immune response, whereas the persistence of HBsAg exposed spleen DC may contribute to long-lasting immunity. This study also suggested that DNA-based vaccines may be a potent tool for treating chronic HBV carriers.

Needle length and injection technique for efficient intramuscular vaccine delivery in infants and children evaluated through an ultrasonographic determination of subcutaneous and muscle layer thickness.

The relationship between resulting reactogenicity and immunogenicity with route and site of vaccine injection is well documented.1-5 Preference for intramuscular injection is given for aluminum-adsorbed vaccines (eg, diphtheria, tetanus, pertussis, and inactivated poliovirus [DTP-IPV], hepatitis A, and hepatitis B vaccines), because superficial administration leads to increased incidence of local reactions.6 A better immune response for intramuscular compared with subcutaneous injection has been seen with several vaccines, such as the hepatitis B,1,5 rabies,3 and influenza2vaccines. Both the injection technique and the needle length are crucial for ensuring proper intramuscular delivery and thus are directly related to vaccine safety and immuno-genicity. Guidelines concerning the choice of the injection technique and needle length have been presented. Two injection techniques are currently recommended. The first, widely used in the United States, requires bunching the thigh muscle at the injection site to increase muscle mass and to minimize the chance of striking bone.6 The second, recommended by the World Health Organization (WHO), suggests stretching the skin flat between the finger and thumb, and pushing the needle down at a 90° angle through the skin.7 With respect to needle length, both the WHO and the Committee on Infectious Diseases of the American Academy of Pediatrics support the use of 7/8-inch (22-mm) or longer needles for intramuscular delivery.7,8 Some unidose vaccines are supplied in disposable syringes, equipped with 5/8-inch (16-mm) sealed needles that have been designed to provide an efficient, precise, and user-friendly tool for intramuscular injection. Nevertheless, the adequacy of this shorter needle (compared with the 7/8-inch needle) has been questioned.6 To determine the optimum needle size for intramuscular injection and eventually to make a correlation between needle length and appropriate injection technique, one must have …

Associations of prevaccination antibody levels with adverse reactions to pneumococcal and influenza vaccines administered simultaneously in the elderly

The association of prevaccination antibodies with the adverse reactions seen after vaccination was studied in 85 elderly subjects (65–90 years) vaccinated simultaneously with pneumococcal and influenza vaccines. The subjects with a temperature rise (9% of vaccinees) had significantly higher prevaccination antibody levels to pneumococcal capsular polysaccharides (PPSs) than those without a temperature rise; no difference was seen in their haemagglutination inhibiting (HI) influenza virus antibody levels. Pain in the left arm (the pneumococcal vaccine injection site) occurred in 45% of the subjects and was likewise associated with elevated PPS antibody levels. Pain at the site of influenza vaccine injection (the right arm) seen in 33% of the vaccinees was significantly more common among those who had previously received influenza vaccine, but was not associated with elevated HI antibody levels. In conclusion, prevaccination pneumococcal but not influenza antibodies were associated with both systemic and local reactions following vaccination.

Tumor vaccines: Effects and fate of IL-2 transfected murine melanoma cells in vivo

We have previously demonstrated the general usefulness of the adenovirus-enhanced transferrinfection (AVET) in the generation of IL-2 producing tumor vaccines. By optimizing different parameters of the transfection protocol we were able to transform the poorly immunogenic M-3 mouse melanoma cell line into a potent immunogen. A long-lasting immunity was demonstrated after administration of the IL-2 releasing vaccine, since immunized animals successfully rejected native M-3 melanoma cells even after a period of more than 6 months. We also demonstrated that in vivo administration of such a vaccine is safe since transmission of the transfected IL-2 gene in host organs was not detected. IL-2 production ceased 2 days after injection because the engineered cells were destroyed. However, RT-PCR analysis of the site of vaccine injection suggests that IL-2 exerts its effects not only directly but also by inducing a set of other immunomodulator cytokines in situ that are probably indispensable in inducing a host response. We conclude that AVET of IL-2 into tumor cells is a safe and efficient method for the generation of tumor vaccines.

SystemicPasteurella HaemolyticaInfection as a Rare Sequel to Avirulent LivePasteurella HaemolyticaVaccination in Cattle

Eleven cases of systemic Pasteurella haemolytica infection in cattle were identified from routine diagnostic laboratory submissions during the falls of 1988, 1989, and 1991. All cases came with a history of recent vaccination with an avirulent live culture P. haemolytica product. Nine of 11 cases involved cattle vaccinated between 2 and 18 days previously with this product. Ten of 11 cases involved 182-227-kg beef calves that were vaccinated between September and November during routine processing for entry into feedlots. The morbidity and mortality was generally low. The major pathologic findings included meningitis, injection site abscessation and/or cellulitis, and polyarthritis. Systemic infection was indicated in all cases by the isolation of P. haemolytica from 2 or more organs or distinct anatomical sites. In 6 cases, the vaccine injection site was cultured, and in all 6 cases, P. haemolytica was isolated. Three separate P. haemolytica isolates from 2 cases were further studied by restriction enzyme analysis (REA). These isolates were from tissues with suppurative inflammation, including the brain, joint, and injection site. The REA patterns of each of these 3 isolates were identical to the REA pattern of the vaccine masterseed, which strongly suggested that the organisms causing systemic infection were the same as the organism used to produce the vaccine. Because the overall incidence was quite low, other factors, such as stress, probably played a major role in the expression of this syndrome.

Postimmunization (Vaccination) Injection-Site Reactions

Four patients each had a single subcutaneous nodule at the site of a previous vaccine injection; three after injection of diphtheria, tetanus, and pertussis vaccination and one after tetanus toxoid vaccination. Presentation was with a mass 4-22 months after vaccination at the site of injection. Histologically, three patients had a necrotizing granulomatous reaction with a surrounding infiltrate of lymphocytes, plasma cells, histiocytes, and associated fibrosis. The fourth patient demonstrated a lymphohistiocytic reaction with a predominance of histiocytic cells as well as associated plasma cells, fibroblasts, and fibrosis. The lymphoid infiltration in these reactions showed a predominance of T-lymphocytes over B-lymphocytes. Aluminum was demonstrated in necrotic foci, inflammatory stroma, and the granular cytoplasm of histiocytes with the aid of solochrome azurine and solochrome cyanine stains as well as by energy-dispersive x-ray microanalysis. The reactions are thought to be immunologic (hypersensitivity) reactions associated with the aluminum contents of the preparation.

Immunisation de chiens avec des vaccins contre la fièvre Q: comparaison entre des vaccins de <em>Coxiella burnetii</em> de phase I, phase II et du RCM de phase I

Q fever vaccines were tested in mixed breed dogs by vaccinating them with formalin-killed Coxiella burnetii whole cells (WC) in either phase I (WCI) or phase II (WCII), or the chloroform: methanol residue (CMR) subunit of phase I cells. Phase I vaccines mixed (1:1) with Freund's incomplete adjuvant (FIA) induced humoral immune responses to phases I and II antigens as measured by microagglutination assay. The CMR vaccine mixed (1:1) with FIA induced greater antigen-specific antibody levels to both phases I and II antigens than the corresponding WCI vaccine. The WCII vaccine induced antibody responses to only phase II antigens. The time course of erythema and induration after skin testing with C. burnetii antigens were suggestive of cell-mediated immunity (CMI). Although granulomas were observed with only WCI and WCII, none of the skin test antigens induced abscesses at the injection site. In contrast, axillary nodes draining the vaccine injection site developed sterile draining abscesses in all dogs by days 19 to 24 for the WCI and CMR, and day 104 for the WCII vaccines. The abscesses had resolved within 30 days after first appearance. Responses to Con A and PHA and recall antigens of lymphocytes from the blood, axillary and mesenteric nodes, and spleen at 222 days after vaccination were variable among dogs. Lymphocytes from various organs responded to one or more of the recall antigens and to both mitogens in the absence or presence of indomethacin. Although these Q fever vaccines induced humoral and CMI, either the antigens or FIA caused sterile draining abscesses.(ABSTRACT TRUNCATED AT 250 WORDS)

Site of injection for vaccination.

Site of injection for vaccination.

Field evaluation of a commercial M-protein vaccine against Streptococcus equi infection in foals

SUMMARY A double-blind randomized clinical trial was undertaken to determine the value of parenterally administered Streptococcus equi M-protein vaccine in foals during an epizootic of strangles. Weaned mixed-breed foals (n = 664) housed on 2 adjacent feed-lots (A and B) arrived over a 5-day period, 2 weeks before primary vaccination. Foals in lot B (n = 114) were randomly administered vaccine (n = 59) or saline solution (placebo; n = 55) on 3 occasions at biweekly intervals. Foals in lot A (n = 450) were given 1 dose of vaccine (n = 225) or placebo. The following clinical observations were scored blindly by a single observer for all foals in lot B and for 120 (randomly sampled) foals in lot A on a single day, 2 (lot B) and 6 (lot A) weeks after final vaccination: cervical lymphadenopathy, type of bilateral nasal discharge, and palpable swelling at injection site(s). Bacteriologic culture of nasal swab specimens or lymph node aspirates from selected foals with clinical disease yielded Sequi. Cervical lymphadenopathy was observed in 17 of 59 (29%) vaccinates and 39 of 55 (71%) nonvaccinated controls in lot B and in 32 of 60 (53%) vaccinates and 29 of 60 (48%) controls in lot A. Contingency χ2analysis confirmed significantly lower cervical lymphadenopathy rate (χ2= 18.5; P < 0.001) and prevalence of mucopurulent nasal discharge (χ2= 11.4; P < 0.01) for vaccinates in lot B only. Swelling(s) at the vaccine injection site were palpated in 44% of lot B and 29% of lot A vaccinates vs < 2% of placebo controls. In the face of intense natural exposure, foals inoculated 3 times with M-protein vaccine were less than half as likely to have clinical signs of strangles as were nonvaccinated horses.

Measles virus panniculitis subsequent to vaccine administration

A severe illness developed gradually in a healthy boy following administration of live measles vaccine at age 14 years. Killed measles vaccine had been given at age 2. Panniculitis began near the site of live vaccine injection, extended contiguously to involve the entire upper arm, and then began to appear in distant subcutaneous sites. He was febrile, leukopenic, anemic, and lost weight rapidly. Evidence of a cause-and-effect relationship between the lesions and the measles vaccine included biopsy findings of particles closely resembling measles nucleocapsids and complete virions by electron microscopy, positive fluorescence after staining with two anti-measles nucleocapsid sera, and immunologic aberrations. The lesions regressed after various treatments and completely remitted on two occasions, but recurred. He became severely malnourished and finally died 29 months after the measles vaccination. At autopsy, pannicultitis was found in the subcutis and mesentery, without evidence of involvement of the brain or other vital organs.

A new approach in specific, active immunotherapy

A vaccinia virus-lysed autochthonous tumor cell vaccine (vaccinia oncolysate) is introduced as a new specific, active immunotherapeutic agent against human cancer. Mouse experiments showed the vaccine to be a safe and potent immune mechanism stimulator. Human experimentation was undertaken in the knowledge of relative safety of the components of the vaccine, i.e. vaccinia vaccine and lysed, autochthonous tumor cells. Vaccine-treated patients had advanced metastatic cancer but reacted to one or more common recall antigen skin tests. None of the 13 patients had untoward responses; 7/13 patients had classic delayed hypersensitivity reactions at the vaccine injection sites; and 2/7 patients with injection site reactions had significant reduction in tumor burden. These results indicate that this vaccine is a specific, active immune mechanism stimulator, and may prove to be a useful therapeutic agent in the treatment of human cancer.

Effect of ionizing radiation on the development of immunity by immunization with live tularemia vaccine

The process of formation of postvaccinal immunity against tularemia after immunization with the living tularemia vaccine is very radioresistant. Experiments were performed on guinea pigs. These animals were irradiated with various x-ray doses, ranging from 10–20 (30) LD to 50–60 (30) LD, at different periods before the vaccination and 24 hr after it. Such irradiation caused only a moderate reduction in intensity of the postvaccinal immunity. The process of postvaccinal immunity formation was found to be even more radioresistant in white mice, in which irradiation provoked an acute exacerbation of the vaccinal infection. The authors compare the ability of the irradiated and normal organisms to form immunity and the intensity of immunobiological reactions (antibody synthesis, inflamuratory reaction at the site of vaccine injection, the ability of the tissues to render the virulentPast.tularensis harmless, allergic reaction to tularin).