Immunization Procedures Research Articles

Monoclonal antibody therapy targeting the shared pathological conformer of both beta-amyloid and hyperphosphorylated tau

Currently there is no effective therapy for Alzheimer's Disease (AD). Active and passive immunomodulation still holds promise but current attempts only address one side of the pathology: either amyloid β (Aβ) or the hyperphosphorylated tau (ptau) protein. Furthermore current approaches are not specific for the pathological conformers of either protein. From our novel immunization method targeting pathological β-sheet conformation (PLoS ONE 5(10): e13391, 2010) we have developed monoclonal antibodies, some of them with promising binding capacities; and we propose to characterize a monoclonal specific for pathological conformers of Aβ and tau (TAB1) to be used for immunotherapy in AD mouse models. Monoclonal antibodies were obtained from our conformational inoculation of BALBc mice. Positive hybridomas were selected by their shared reactivity against Aβ, PHF and PrP Res. The best conformational candidate (TAB1) was selected after characterization by blots, ELISA and histology against pathological conformers in AD tissue. TAB1 is being used of treatment by intraperitoneal injection in 3xTg mice with both tau and Aβ related pathology. TAB1 on tissue sections specifically immunolabels AD tissue with no labeling in young, health normal control brains. On Western blots TAB1 detects purified paired helical filament preparations, aggregated/oligomeric Aβ and PrP Res, extracted from CJD brain tissue. Use of TAB1 therapeutically in 3xTg mice is on-going. We have developed a novel immunization procedure which we have used to produce monoclonal antibodies (mAbs) that recognize multiple pathological proteins, including PrP Res, oligomer A b and ptau. We have characterized one of these mAbs, TAB1, which gives specific immunolabeling in AD tissue and on Western blots to pathological conformers. We believe that immunotherapy that specifically targets the most toxic, oligomeric forms of Aβ and ptau, has the greatest chance of success with little risk of toxicity.

Instance selection for time series classification based on immune binary particle swarm optimization

We propose a new immune binary particle swarm optimization algorithm (IBPSO) to solve the problem of instance selection for time series classification, whose objective is to find out the smallest instance combination with maximal classification accuracy. The proposed IBPSO is based on the basic binary particle swarm optimization (BPSO) algorithm proposed by Kennedy and Eberhart. Its immune mechanism includes vaccination and immune selection. Vaccination employs the hubness score of time series and the particles’ inertance as heuristic information to direct the search process. Immune selection procedure always discards the particle with the worst fitness in the current swarm for preventing the degradation of the swarm. Experimental results on small and medium datasets show that IBPSO outperforms BPSO and deterministic INSIGHT in terms of storage requirement and classification accuracy, and presents better robustness to noise than BPSO. In addition, experimental results on larger datasets indicate that IBPSO has better scalability than BPSO.

Development of claudin‐1‐specific ligand

Claudins (CLs) are a family of tetratransmembrane proteins consisting of 27 members and are potential targets for enhancing mucosal drug absorption, treating cancer, and mucosal vaccination using the C‐terminal fragment of Clostridium perfringens enterotoxin (C‐CPE), a CL‐3/‐4 binder. The development of efficient CL binders is important for their future clinical application. We previously developed a broadly specific CL binder by using C‐CPE as a prototype; however, we did not successfully generate a CL subtype‐specific binder. Here, we focused on developing a CL subtype‐specific antibody. Because of the low antigenicity of CLs, we performed three different immunization procedures. BXSB mice with an autoimmune disorder were immunized with CL‐1‐displayed budded baculovirus, CL‐1‐expressing cells, or plasmid DNA encoding CL‐1. The DNA‐based immunization showed the highest production of serum anti‐CL‐1 antibodies. Therefore, B cells were isolated from the DNA‐immunized mice, and the B cells were fused with myeloma cells to form hybridomas. Hybridomas were screened for the production of the anti‐CL‐1 antibody. The antibodies bound to CL‐1‐expressing cells, but not to CL‐2, ‐4 or ‐5‐expressing cells. These results indicate that immunization of BXSB mice with CL DNA is a potentially useful method for the development of specific CL ligands.

Characterization of scFv libraries derived from mice differently immunized with claudin

Claudins (CLs), a family of tetratransmembrane proteins consisting of 27 members, are potential targets for enhancing drug absorption, treating cancer, and vaccination. The small extracellular loop regions of CL proteins have made it difficult to develop CL ligands, including antibodies, and to generate recombinant CL proteins, except for CL‐4. Here, we optimized immunization procedures for developing antibody phage display libraries in a single‐chain antibody fragment (scFv) format. CL‐1‐displaying budded baculovirus (CL‐1‐BV) and plasmid DNA encoding the CL‐1 gene were the antigens. BXSB mice, which spontaneously develop a lupus‐like syndrome, were immunized with CL‐1‐BV. The scFv library comprised 7.95 × 105 colony formation units (CFU); however, most clones bound to both mock‐BV and CL‐1‐BV, indicating that the clones may recognize gp64 derived from BV. To overcome this immune response, gp64 transgenic mice were immunized with CL‐1‐BV and used to generate a scFv library up to 2.6 × 105 CFU. We isolated 6 phage clones that bound to CL‐1, ‐2, ‐4 and ‐5, but not to mock‐BV. Furthermore, we got a specific CL‐1‐bound clone. We also prepared a scFv library up to 1.1 × 106 CFU from mice immunized with CL‐1 DNA, but a phage clone that bound to CL‐1, ‐2, ‐4 and ‐5 was isolated. These findings indicate that the gp64 library might be the most promising library for preparing CL‐binding scFv fragments.

The enigma of IgE+ B-cell memory in human subjects

Our understanding of the origin and fate of the IgE-switched Bcell has been markedly improved by studies in mouse models. The immediate precursor of the IgE-switched B cell is either a relatively naive nonswitched B cell or a mature IgG-switched Bcell. These 2 routes are referred to as the direct and indirect pathways, respectively. IgE responses derived from each pathway differ significantly, largely reflecting the difference in time spent in a germinal center and thus time for clonal expansion, somatic hypermutation, affinity maturation, and acquisition of a memory phenotype. The clinical and therapeutic implications for IgE responses in human subjects are still a matter of debate, largely because the immunization procedures used in the animal models are significantly different from classical atopic sensitization to allergens from pollen and mites. On the basis of the limited information available, it seems likely that these atopic IgE responses are characterized by a relatively low IgG/IgE ratio, low B-cell memory, and modest affinity maturation, which fits well with the direct switching pathway. It is still unresolved how the IgE response evolves to cover a wide epitope repertoire involving many epitopes per allergen, as well as many different allergens from a single allergen source.

The Twenty-Year Story of a Plant-Based Vaccine Against Hepatitis B: Stagnation or Promising Prospects?

Hepatitis B persists as a common human disease despite effective vaccines having been employed for almost 30 years. Plants were considered as alternative sources of vaccines, to be mainly orally administered. Despite 20-year attempts, no real anti-HBV plant-based vaccine has been developed. Immunization trials, based on ingestion of raw plant tissue and conjugated with injection or exclusively oral administration of lyophilized tissue, were either impractical or insufficient due to oral tolerance acquisition. Plant-produced purified HBV antigens were highly immunogenic when injected, but their yields were initially insufficient for practical purposes. However, knowledge and technology have progressed, hence new plant-derived anti-HBV vaccines can be proposed today. All HBV antigens can be efficiently produced in stable or transient expression systems. Processing of injection vaccines has been developed and needs only to be successfully completed. Purified antigens can be used for injection in an equivalent manner to the present commercial vaccines. Although oral vaccines require improvement, plant tissue, lyophilized or extracted and converted into tablets, etc., may serve as a boosting vaccine. Preliminary data indicate also that both vaccines can be combined in an effective parenteral-oral immunization procedure. A partial substitution of injection vaccines with oral formulations still offers good prospects for economically viable and efficacious anti-HBV plant-based vaccines.

Establishing Intra‐ and Inter‐Rater Agreement of the Face, Legs, Activity, Cry, Consolability Scale for Evaluating Pain in Toddlers During Immunization

The Face, Legs, Activity, Cry, Consolability (FLACC) scale is a five-item tool that was developed to assess postoperative pain in young children. The tool is frequently used as an outcome measure in studies investigating acute procedural pain in young children; however, there are limited published psychometric data in this context. To establish inter-rater and intrarater agreement of the FLACC scale in toddlers during immunization. Participants comprised a convenience sample of toddlers recruited from an immunization drop-in service, who were part of a larger pilot randomized controlled trial. Toddlers were video- and audiotaped during immunization procedures. The first rater scored each video twice in random order over a period of three weeks (intrarater agreement), while the second rater scored each video once and was blinded to the first rater's scores (inter-rater agreement). The FLACC scale was scored at four timepoints throughout the procedure. Intraclass correlation coefficients were used to assess agreement of the FLACC scale. Thirty toddlers between 12 and 18 months of age were recruited, and video data were available for 29. Intrarater agreement coefficients were 0.88 at baseline, 0.97 at insertion of first needle, and 0.80 and 0.81 at 15 s and 30 s following the final injection, respectively. Inter-rater coefficients were 0.40 at baseline, 0.95 at insertion of first needle, and 0.81 and 0.78 at 15 s and 30 s following the final injection, respectively. The FLACC scale has sufficient agreement in assessing pain in toddlers during immunizations, especially during the most painful periods of the procedure.

Evaluation of Pre-Clinical Efficacy to HPV16 L2E6E7 Vaccine and HPV16 E6E7 Adenovirus-5 Vector Vaccine with Different Dosages and Prime- Booster Regiments in Mouse Model

Purpose: This study evaluates the dose-response and immunization procedure in mouse model and the efficacy of prime-booster regimens with HPV16 L2E6E7 vaccine and HPV16 E6E7 Ad5 vector vaccine. Methods: Experimental animals were C57 BL/6 mice. Each group included 10 or 20 C57 BL/6 mice. The tumor model used TC-1 tumor cells. The HPV16 L2E6E7 vaccine groups were treated using the following dosage: 15 μg/ml, 30 μg/ml, 60 μg/ml, 120 μg/ml, 240 μg/ml, and then 120 μg/ml was used for the following regimens: 0-7 days, 0-15 days, 0-7-15 days. The HPV16 E6E7 Adenovirus-5 vector vaccine groups were treated using the following dosage: 3.00×106 IU/ml, 3.00×107 IU/ml, 3.00×108 IU/ml, 3.00×109 IU/ml, and then 3.00×107 IU/ml was used for the following regimens: 0-7 days, 0-15 days, 0-7-15 days, and control group. Prime-booster combined regimens with HPV L2E6E7 vaccine (P, 120 μg/ml) and HPV16 E6E7 ad5 vector vaccine (V, 3.00×107 IU/ml) were set as follows: 0P-7P days, 0P-7V days, and 0P-7P-15V days, 0P-7V-15V days, and 0P-7P-15V-21V days. Results: Upon challenge with 104 TC-1 tumor cells, mice developed palpable, rapidly growing tumors within 7–14 days. These tumors became lethal to the mice within 21–28 days. HPV16 L2E6E7 vaccine (120 μg/ml, 0-7- 15 day’s procedure) protective efficacy was 85% and the HPV16 E6E7 Ad5 vector vaccine (3.00×107 IU/ml, 0 day procedure) that was 80%. Prime-booster regimens showed a protective efficacy of 80–90% for the 0P-7V days and 0P-7V-15V day’s schedules. Conclusion: HPV16 L2E6E7 vaccine and HPV16 E6E7 Ad5 vector vaccine are proved the candidate vaccine for therapeutic intervention against HPV16-induced tumor.

Secretory IgA: Designed for Anti-Microbial Defense.

Prevention of infections by vaccination remains a compelling goal to improve public health. Mucosal vaccines would make immunization procedures easier, be better suited for mass administration, and most efficiently induce immune exclusion – a term coined for non-inflammatory antibody shielding of internal body surfaces, mediated principally by secretory immunoglobulin A (SIgA). The exported antibodies are polymeric, mainly IgA dimers (pIgA), produced by local plasma cells (PCs) stimulated by antigens that target the mucose. SIgA was early shown to be complexed with an epithelial glycoprotein – the secretory component (SC). A common SC-dependent transport mechanism for pIgA and pentameric IgM was then proposed, implying that membrane SC acts as a receptor, now usually called the polymeric Ig receptor (pIgR). From the basolateral surface, pIg-pIgR complexes are taken up by endocytosis and then extruded into the lumen after apical cleavage of the receptor – bound SC having stabilizing and innate functions in the secretory antibodies. Mice deficient for pIgR show that this is the only receptor responsible for epithelial export of IgA and IgM. These knockout mice show a variety of defects in their mucosal defense and changes in their intestinal microbiota. In the gut, induction of B-cells occurs in gut-associated lymphoid tissue, particularly the Peyer’s patches and isolated lymphoid follicles, but also in mesenteric lymph nodes. PC differentiation is accomplished in the lamina propria to which the activated memory/effector B-cells home. The airways also receive such cells from nasopharynx-associated lymphoid tissue but by different homing receptors. This compartmentalization is a challenge for mucosal vaccination, as are the mechanisms used by the mucosal immune system to discriminate between commensal symbionts (mutualism), pathobionts, and overt pathogens (elimination).

Long-term response rates of successful hepatitis B vaccination in HIV-infected patients

BackgroundData on long-term response rates after successful primary hepatitis B (HBV) vaccination in HIV-infected patients are scarce. ObjectiveTo evaluate the durability of an effective anti-HBs titer up to 5 years after primary vaccination in a cohort of 155 HIV-infected adults. MethodsFrom a previous multicenter HBV vaccination trial we selected patients with an anti-HBs titer of ≥10IU/l 28 weeks after the first vaccination. The anti-HBs titer was measured in annually stored plasma samples up to 5 years after vaccination. Patients with decreasing anti-HBs titers <10IU/I were defined as transient responders (TR*) and with persistent anti-HBs titers ≥10IU/I as long-term responders (LTR^). ResultsWe included 155 patients, 87 were TR and 68 LTR. Mean age, percentage of female participants and duration of HAART use at primary vaccination were similar in LTR and TR. Anti-HBs level after primary vaccination was the strongest predictor for the durability of anti-HBs. Anti-HBs >100–1000IU/I and >1000 resulted in an OR 8.3, 95% CI 3.38–20.16; p<0.0001 and OR 75.6, 95% CI 13.41–426.45; p<0.0001 versus anti-HBs titer of 10–100IU/I after primary vaccination respectively. The mean time to loss of an effective anti-HBs titer was 2.0, 3.7 and 4.4 years respectively, for patients with an anti-HBs titer of 10–100IU/I, >100–1000IU/I and >1000IU/I at primary vaccination. An undetectable HIV-RNA load and use of HAART during vaccination and at follow-up were, though not significantly, associated a higher long-term persistence of an effective antibody titer. ConclusionThe durability of an effective anti-HBs level appears to be significantly related to the height of the antibody titers after the primary immunization procedure. Schedules to improve the vaccination response in HIV-infected patients therefore seem to be justified. Whether a HBV booster is indicated remains to be elucidated.

Formatted single-domain antibodies can protect mice against infection with influenza virus (H5N2)

This work continues a series of recently published studies that employ recombinant single-domain antibody (sdAb, or nanobody®) generation technologies to battle viruses by a passive immunization approach. As a proof of principle, we describe a modified technique to efficiently generate protective molecules against a particular strain of influenza virus within a reasonably short period of time. This approach starts with the immunization of a camel (Camelus bactrianus) with the specified antigen-enriched material presented in as natural a form as possible. An avian influenza virus A/Mallard/Pennsylvania/10218/84 (H5N2) adapted for mice was used as a model source of antigens for both the immunization and phage display-based selection procedures. To significantly increase activities of initially selected monovalent single-domain antibodies, we propose a new type of sdAb formatting that involves the addition of a special type of coiled-coil sequence, the isoleucine zipper domain (ILZ). Presumably, the ILZ-containing peptides adopt trimeric parallel conformations. After the formatting, the biological activities (virus neutralization) of the initially selected anti-influenza virus (H5N2) sdAbs were significantly increased. Intraperitoneal or intranasal administration of the formatted sdAb at 2h before or 24h after viral challenge specifically protects mice from lethal infection with influenza virus. We hope that the described approach combined with the selection focused on particular conservative epitopes will lead to the generation of sdAb-based molecules protective against a broad spectrum of influenza virus subtypes.

Robust Immunity and Heterologous Protection against Influenza in Mice Elicited by a Novel Recombinant NP-M2e Fusion Protein Expressed in E. coli

BackgroundThe 23-amino acid extracellular domain of matrix 2 protein (M2e) and the internal nucleoprotein (NP) of influenza are highly conserved among viruses and thus are promising candidate antigens for the development of a universal influenza vaccine. Various M2e- or NP-based DNA or viral vector vaccines have been shown to have high immunogenicity; however, high cost, complicated immunization procedures, and vector-specific antibody responses have restricted their applications. Immunization with an NP–M2e fusion protein expressed in Escherichia coli may represent an alternative strategy for the development of a universal influenza vaccine.Methodology/Principal FindingscDNA encoding M2e was fused to the 3′ end of NP cDNA from influenza virus A/Beijing/30/95 (H3N2). The fusion protein (NM2e) was expressed in E. coli and isolated with 90% purity. Mice were immunized with recombinant NM2e protein along with aluminum hydroxide gel and/or CpG as adjuvant. NM2e plus aluminum hydroxide gel almost completely protected the mice against a lethal (20 LD50) challenge of heterologous influenza virus A/PR/8/34.Conclusions/SignificanceThe NM2e fusion protein expressed in E. coli was highly immunogenic in mice. Immunization with NM2e formulated with aluminum hydroxide gel protected mice against a lethal dose of a heterologous influenza virus. Vaccination with recombinant NM2e fusion protein is a promising strategy for the development of a universal influenza vaccine.

Is An Oral Plant-based Vaccine against Hepatitis B Virus Possible?

Prevention of hepatitis B, one of the most prevalent human diseases, still requires cheap and commonly available vaccines. Oral vaccines, including plant-based formulations, have been considered as alternatives or supplements for standard injection vaccines, due to the assumed low-cost production and simplified vaccination. Although plant production of HBV antigens is sufficiently efficient, despite almost 20 years of research still no anti-HBV plant-based vaccine has been developed. The basic difficulty has been to elaborate an effective immunisation procedure. Immunisation by parenteral priming and oral boosting with raw plant tissue adjuvanted with the cholera toxin, although effective, seemed to be unfeasible and controversial. Exclusively oral immunisation using lyophilised tissue, despite its appropriate form, appeared also impractical because of too low efficiency. Oral tolerance turned out to be the main barrier for anti-HBV plant-based vaccines. Based on previous results and knowledge on the mucosal immune system, a possible vaccine may consist of two components, parenteral for priming and oral for boosting. Probably the oral constituent could independently serve for further booster vaccinations. Both vaccine components can be produced in plants and used after some processing - purification for the injection constituent and lyophilisation for the oral one. Lyophilised tissue can be converted into tablets, capsules, etc. Previous and recent data show that the injection-oral immunisation regime may be efficient. A combination of parenteral and oral vaccination offers good prospects for a truly efficacious plant-derived anti-HBs vaccine and even a partial substitution of parenteral vaccines by an oral formula may prove to be economically reasonable.

Murine model of Graves disease induced by thyroid-stimulating hormone receptor gene transfected via liposome

To compare the efficacy of different expression vectors, target genes, and immunization procedures in transfecting mice via liposome to construct murine model of Graves disease. We linked pCDNA3.1(+) and pUBC to full-length human TSHR and TSHR A subunit cDNA to yield four plasmids, which were later injected intramascularly or subcutaneously into female Balb/c mice via liposome. The blood anti-TSHR antibody (TRAb) were determined and the body weight were measured after each immunization. Serum thyroid hormone levels were measured after the animals were sacrificed. In mice immunized with pUBC, no significant variance with control in weight nor serum TRAb concentration was observed. Weight gain in pCDNA3.1(+) group was significantlyly slower than controls (p<0.05), and serum TRAb concentration was also significantly elevated. In pCDNA group, animals immunized with TSHR A subunit (TSHRA subgroup) as the target gene revealed even significantly slower weight gain (p<0.001) and even faster TRAb elevation than those immunized with full length TSHR. Significantly higher FT4 (p=0.023) was observed in TSHRA and TSHR subgroups, which was reversely correlated to weight gain, but no significant difference (p>0.05) in FT3 was observed. Weight gain and TRAb concentration mainly varied in the later period of immunization. Immunization with pCDNA3.1(+) and TSHR A subunit gene together with higher immunization frequency increases the chance of model induction. Furthermore, FT4 is a better indicator for assessing the thyroid function in this model.

HIV infection

At a satellite meeting prior to the International AIDS Meeting in Washington DC in July, a great deal of discussion was focused on finding a “cure” for HIV infection [1]. While this objective has been discussed at various times over the past decade, the real incentive for this goal comes from the encouraging findings with Timothy Brown, also known as the “Berlin patient.” This HIV-infected man received hematopoietic stem cells from an HLA-matched donor whose genome naturally lacked CCR5, the major receptor for HIV. Five years after two transplants, Mr. Brown appears to be cured of HIV as well as leukemia [2]. Although potentially infected with both a virus that requires CCR5 for entry and a small population of viruses that can infect cells by another receptor, no infectious virus can be recovered from his blood and other tissues. This finding includes biopsies from the brain, bowel, and lymph nodes. Certainly his “cure” encourages approaches that could mimic this achievement in all HIV-infected individuals.

Nasal immunization with major epitope-containing ApxIIA toxin fragment induces protective immunity against challenge infection with Actinobacillus pleuropneumoniae in a murine model

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439–801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.

Oral sucrose for pain control in nonneonate infants during minor painful procedures

To provide information regarding the effective use of oral sucrose as an analgesic for immunization and venipuncture procedures in the older infant. Evidence-based literature including original clinical trials, reviews, and clinical practice guidelines. Most infants are exposed to multiple minor painful procedures during the first year of life. Oral sucrose solution in a 24% concentration at a dose of 2 mL approximately 2 min prior to the painful procedure has been shown effective in reducing pain during immunizations and venipuncture in the outpatient setting in infants aged 1-12 months old. Oral sucrose solution should be used as a pain reduction intervention in infants up to 12 months of age undergoing minor painful procedures. Its proven effectiveness as an analgesic, low rate of minor adverse events, ease of administration, and excellent availability make sucrose a good choice for this purpose. Additional research is needed regarding dose response in different infant age groups, optimal concentration of sucrose solution, need for multiple sucrose dosing, adjustment for multiple painful procedures, and addition of optimal nonpharmacologic interventions as a combination approach.

Effects of maternal parity, depression and stress on two-month-old infant expression of pain

Objective: This study investigates effects of maternal parity, mental health and frequency of touch on infant distress expressed during routine vaccination, as well as the ability of mothers to assess infant pain. Methods: 50 mothers (19 first-time mothers, 23 stressed, 8 depressed) and their 2-month-old infants (mean age 8.1 weeks, range 7–9 weeks) were videotaped during routine immunisation for off-line-analysis. Infant- related pain behaviours were coded before, during and after needle insertion. Maternal touch was coded throughout the immunisation procedure and maternal estimates of infant pain were established immediately after the procedure. Results: Infants of first-time mothers showed significantly more pain before the needle insertion as well as during the first vaccination, compared with infants of experienced mothers. In contrast to expectation, maternal mental health status and type of touch did not influence the level of pain expressed by infants. Mothers consistently overestimated infant pain and their assessments were poorly correlated with observed infant pain behaviours. Conclusion: These findings suggest that maternal parity may influence infant pain expression before vaccination. However, all mothers overestimate their infant’s pain. Further studies are required to resolve conflicting findings in this field of research, as findings may have policy implications in promoting adherence to the vaccination schedule.

Vaccine allergy evaluation and management at the specialized Green Channel Consultation Clinic

Suspected vaccine allergy may be a cause of incomplete or delayed vaccination. Patients at risk of adverse reactions or suspected contraindications need specialized consultation about subsequent vaccinations. To analyse consultancy results for patients at risk of allergic reactions to vaccines as evaluated by the Green Channel University Hospital Immunization Consultancy Clinic. A review of cases of allergic reactions to vaccines or contraindications due to underlying diseases or sensitization to vaccine components submitted to the Green Channel was carried out. Analysed data included detailed clinical reaction history, skin and in vitro allergy testing with vaccine components, recommendations for vaccination and outcome of subsequent vaccine administrations. A total of 519 cases, 370 referred for previous local or systemic reactions to vaccines, mostly cutaneous, and 149 sent for suspected contraindications were evaluated. Skin testing was performed on 152 patients, specific IgE determination in 37 subjects and patch testing in 173 cases. After consultation, 442 (85%) subjects were advised to continue vaccination, with personalized precautions (premedication, or alternative brand, or administration in graded doses) for 200 of them. Among the 352 (80%) patients vaccinated as per Green Channel instructions, 33 subjects (9.3%) reported mild allergic or non-specific symptoms and one (0.3%) urticaria with bronchospasm. Even though vaccine allergy occurs very rarely, a safe procedure for immunization can be applied, through specialized allergy consultancy, for most subjects with suspected allergy to vaccines, and who could be potentially excluded from vaccination for risk of adverse reactions.

DNA vaccine encoding α-fetoprotein fused with the ornithine decarboxylase degradation signal significantly suppresses the hepatocellular carcinoma growth in mice

Hepatocellular carcinoma (HCC) is the most frequent primary malignancy of the liver in humans. The available anti-HCC therapies are ineffective, and their use has substantial limitations. DNA vaccines expressing α-fetoprotein (AFP), which is a marker of HCC, were recently found to induce the immune response against AFP in vivo. However, the vectors used as vaccines failed to ensure the necessary level of protection, and the immunization procedures were rather complex. We showed earlier that a DNA vaccine encoding HIV-1 reverse transcriptase (RT) with the C end fused with the ornithine decarboxylase (ODC) degradation signal induced a strong Th-1 immune response against RT in mice. In this work, we designed a set of novel DNA vaccines encoding AFP or AFP with the ODC degradation signal. Transfected cells demonstrated efficient expression and fast proteasomal degradation of the recombinant AFP. The antitumor immune response was stimulated in immunized animals, and a notable retardation of HCC growth was observed as a result of protective vaccination.